U.S. patent application number 15/017488 was filed with the patent office on 2016-08-11 for systems and methods for assessing biological samples.
The applicant listed for this patent is LIFE TECHNOLOGIES CORPORATION. Invention is credited to Mauro AGUANNO, Kuan Moon BOO, Mingsong CHEN, Soo Yong LAU, Wei Fuh TEO, Tiong Han TOH, Huei Steven YEO.
Application Number | 20160230210 15/017488 |
Document ID | / |
Family ID | 55404853 |
Filed Date | 2016-08-11 |
United States Patent
Application |
20160230210 |
Kind Code |
A1 |
CHEN; Mingsong ; et
al. |
August 11, 2016 |
SYSTEMS AND METHODS FOR ASSESSING BIOLOGICAL SAMPLES
Abstract
An instrument for biological analysis includes a base, an
excitation source, an optical sensor, an excitation optical system,
and an emission optical system. The base is configured to receive a
sample holder comprising a plurality of biological samples. The
optical sensor is configured to receive emissions from the
biological samples in response to the excitation source. The
instrument may additionally include a sensor lens enclosed by a
lens case and a focusing mechanism including a gear that engages
the lens case, the focusing mechanism being accessible outside the
enclosure for adjusting a focus. The may instrument further include
a sensor aperture dispose along an emission optical path and a
blocking structure disposed to cooperate with the sensor aperture
such that none of the reflected radiation from an illuminated
surface near the sample holder is received by the optical sensor
that does not also reflect off another surface of the
instrument.
Inventors: |
CHEN; Mingsong; (Singapore,
SG) ; BOO; Kuan Moon; (Singapore, SG) ; TOH;
Tiong Han; (Singapore, SG) ; AGUANNO; Mauro;
(Singapore, SG) ; LAU; Soo Yong; (Singapore,
SG) ; YEO; Huei Steven; (Singapore, SG) ; TEO;
Wei Fuh; (Singapore, SG) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
LIFE TECHNOLOGIES CORPORATION |
Carlsbad |
CA |
US |
|
|
Family ID: |
55404853 |
Appl. No.: |
15/017488 |
Filed: |
February 5, 2016 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
62112910 |
Feb 6, 2015 |
|
|
|
Current U.S.
Class: |
1/1 |
Current CPC
Class: |
B01L 2300/168 20130101;
G01N 21/6458 20130101; B01L 2300/18 20130101; G01N 2021/6463
20130101; G01N 21/6428 20130101; C12Q 1/686 20130101; B01L
2300/0654 20130101; G01N 21/6452 20130101; B01L 2200/10 20130101;
G02B 21/244 20130101; B01L 7/52 20130101; G01N 21/6456 20130101;
B01L 2300/0829 20130101 |
International
Class: |
C12Q 1/68 20060101
C12Q001/68; B01L 7/00 20060101 B01L007/00; G01N 21/64 20060101
G01N021/64 |
Claims
1. An instrument for biological analysis, comprising: an excitation
source; an optical sensor configured to receive emissions from a
biological samples in response to the excitation source; an
excitation optical system disposed along an excitation optical
path; an emission optical system disposed along an emission optical
path; an imaging unit comprising: a cavity comprising a first
surface, an opposing second surface, and a threaded housing; a
sensor lens; and a focusing mechanism comprising a focusing gear
configured to engage the threaded housing so as to move the sensor
lens along an optical axis thereof; wherein the first surface
comprises a surface of the sensor lens; wherein the second surface
comprises an optical sensor circuit board; and wherein the sensor
lens is adjustable by indirect engagement via the focusing
gear.
2. An instrument for biological analysis, comprising: an excitation
source; an optical sensor configured to receive emissions from the
biological samples in response to the excitation source; an
excitation optical system disposed along an excitation optical
path; an emission optical system disposed along an emission optical
path; a sensor lens configured to direct emissions from at least
some of the biological sample onto the optical sensor; an
illuminated surface disposed along the excitation optical path, the
illuminated surface configured to produce reflected radiation
comprising radiation from the excitation source that is reflected
by the illuminated surface; a radiation shield, comprising: a
sensor aperture dispose along the emission optical path; and a
blocking structure configured to cooperate with the sensor aperture
such that none of the reflected radiation is received by the
optical sensor that does not also reflect off another surface of
the instrument.
3. An instrument for biological analysis, comprising: an excitation
source; an optical sensor configured to receive emissions from the
biological samples in response to the excitation source; an
excitation optical system disposed along an excitation optical
path; an emission optical system disposed along an emission optical
path; an energy or power detection unit comprising: an energy or
power sensor located outside the optical paths; and a light pipe
disposed adjacent the a a beamsplitter and configured to transport
radiation from the beamsplitter to the power sensor.
4-6. (canceled)
7. An instrument according to claim 1 further comprising a
beamsplitter disposed along both the excitation optical path and
along the emission optical path.
8-9. (canceled)
10. The instrument according to claim 7, further comprising: a base
configured to receive a sample holder comprising a plurality of
spatially separated reaction regions for processing one or more
biological samples; wherein the excitation source is configured to
produce a first excitation beam characterized by a first wavelength
and a second excitation beam characterized by a second wavelength
that is different from the first wavelength.
11-16. (canceled)
17. The instrument according to claim 1, wherein at least some of
the emissions comprise a fluorescent emission from at least some of
the biological samples in response to at least one of the
excitation beams.
18-21. (canceled)
22. The instrument according to claim 7, further comprising a
mirror disposed along the excitation optical path between the base
and the beamsplitter.
23. The instrument according to claim 10, wherein the base
comprises a sample block assembly configured to control the
temperature of the sample holder or biological samples.
24-26. (canceled)
27. The instrument according to claim 10, wherein the base
comprises a thermal cycler configured to perform a PCR assay.
28-32. (canceled)
33. The instrument according to claim 10, wherein the sample holder
comprises a microtiter plate and the reaction regions comprise at
least 96 well, at least 384, or at least 1536 wells.
34-37. (canceled)
38. The instrument according to claim 1, wherein the excitation
optical system comprises a sample lens configured to direct the
excitation beams toward the base.
39. The instrument according to claim 38, wherein the sample lens
comprises a field lens configured to extend over the plurality of
spatially separated regions.
40-43. (canceled)
44. The instrument according to claim 38, wherein the sample lens
comprises a plurality of lenses corresponding to the plurality of
reaction regions.
45-49. (canceled)
50. The instrument according to claim 7, wherein the beamsplitter
comprises a 50/50 beamsplitter.
51-54. (canceled)
55. The instrument according to claim 1, wherein the excitation
light source comprises a light emitting diode.
56-70. (canceled)
71. The instrument according to claim 68, wherein the excitation
filters comprise a plurality of filters together providing a
plurality of band passes suitable for fluorescing one or more of a
SYBR.RTM. dye or probe, a FAM.TM. dye or probe, a VIC.RTM. dye or
probe, a ROX.TM. dye or probe, or a TAMRA.TM. dye or probe.
72. The instrument according to claim 68, wherein the excitation
filters are mounted onto a rotatable filter wheel configure to move
each of the filters into and out of the excitation beam path.
73. The instrument according to claim 1, wherein the excitation
source comprises a plurality of individual excitation sources.
74-81. (canceled)
82. The instrument according to claim 10, further comprising a
heated cover disposed adjacent the base and including a plurality
of apertures configured to correspond to the plurality of reaction
regions.
83. The instrument according to claim 7, further comprising a 50/50
beamsplitter, wherein the beamsplitter: passes the first excitation
beam and the second excitation beam; and passes the first emission
beam and the second emission beam.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application claims the benefit of priority of U.S.
Provisional Patent Application No. 62/112,910, filed on Feb. 6,
2015, which is incorporated herein in its entirety by
reference.
BACKGROUND OF THE INVENTION
[0002] 1. Field of the Invention
[0003] The present invention relates generally to systems, devices,
and methods for observing, testing, and/or analyzing one or more
biological samples, and more specifically to systems, devices, and
methods comprising an optical system for observing, testing, and/or
analyzing one or more biological samples.
[0004] 2. Description of the Related Art
BRIEF DESCRIPTION OF THE DRAWINGS
[0005] Embodiments of the present invention may be better
understood from the following detailed description when read in
conjunction with the accompanying drawings. Such embodiments, which
are for illustrative purposes only, depict novel and non-obvious
aspects of the invention. The drawings include the following
figures:
[0006] FIG. 1 is a schematic representation of a system according
to an embodiment of the present invention.
[0007] FIG. 2 is a schematic representation of an excitation source
according to an embodiment of the present invention.
[0008] FIG. 3 is a normalized spectrum plot of various light
sources, including a light source according to an embodiment of the
present invention.
[0009] FIG. 4 is plot of spectral integration over various
wavelength ranges for the light source spectrums shown in FIG.
3
[0010] FIGS. 5 and 6 are perspective views of an instrument housing
according to an embodiment of the present invention.
[0011] FIG. 7 is a solid model representation of an optical and
sample processing system according to an embodiment of the present
invention.
[0012] FIG. 8 is a magnified, solid model representation of the
optical system shown in FIG. 7.
[0013] FIG. 9 is an exploded view of a portion of the sample
processing system shown in FIG. 7.
[0014] FIG. 10 is a section view of a portion of the optical system
shown in FIG. 7.
[0015] FIG. 11 is a top perspective view of an imaging unit
according to an embodiment of the present invention.
[0016] FIG. 12 is a sectional view of the imaging unit shown in
FIG. 11
[0017] FIGS. 13 and 14 are bottom perspective views of the imaging
unit shown in FIG. 11.
[0018] FIGS. 15-17 are magnified views of portions of the imaging
unit shown in FIG. 11.
[0019] FIG. 18 is a section view of the system shown in FIGS. 6 and
8.
[0020] FIGS. 19 and 20 are schematic representations of a system
according to an embodiment of the present invention.
DETAILED DESCRIPTION OF THE DRAWINGS
[0021] As used herein the terms "radiation" or "electromagnetic
radiation" means radiant energy released by certain electromagnetic
processes that may include one or more of visible light (e.g.,
radiant energy characterized by one or more wavelengths between 400
nanometers and 700 nanometers or between 380 nanometers and 800
nanometers) or invisible electromagnetic radiations (e.g.,
infrared, near infrared, ultraviolet (UV), X-ray, or gamma ray
radiation).
[0022] As used herein an excitation source means a source of
electromagnetic radiation that may be directed toward at least one
sample containing one or more chemical compounds such that the
electromagnetic radiation interacts with the at least one sample to
produce emission electromagnetic radiation indicative of a
condition of the at least one sample. The excitation source may
comprise light source. As used herein, the term "light source"
refers to a source of electromagnetic radiation comprising an
electromagnetic spectrum having a peak or maximum output (e.g.,
power, energy, or intensity) that is within the visible wavelength
band of the electromagnetic spectrum (e.g., electromagnetic
radiation within a wavelength in the range of 400 nanometers to 700
nanometers or in the range of 380 nanometers and 800 nanometers).
Additionally or alternatively, the excitation source may comprise
electromagnetic radiation within at least a portion of the infrared
(near infrared, mid infrared, and/or far infrared) or ultraviolet
(near ultraviolet and/or extreme ultraviolet) portions of the
electromagnetic spectrum. Additionally or alternatively, the
excitation source may comprise electromagnetic radiation in other
wavelength bands of the electromagnetic spectrum, for example, in
the X-ray and/or radio wave portions of the electromagnetic
spectrum. The excitation source may comprise a single source of
light, for example, an incandescent lamp, a gas discharge lamp
(e.g., Halogen lamp, Xenon lamp, Argon lamp, Krypton lamp, etc.), a
light emitting diode (LED), an organic LED (OLED), a laser, or the
like. The excitation source may comprise a plurality of individual
light sources (e.g., a plurality of LEDs or lasers). The excitation
source may also include one or more excitation filters, such as a
high-pass filter, a low-pass filter, or a band-pass filter. For
example, the excitation filter may include a colored filter and/or
a dichroic filter. The excitation source comprise a single beam or
a plurality of beams that are spatially and/or temporally
separated.
[0023] As used herein, an "emission" means an electromagnetic
radiation produced as the result an interaction of radiation from
an excitation source with one or more samples containing, or
thought to contain, one or more chemical and/or biological
molecules or compounds of interest. The emission may be due to a
reflection, refraction, polarization, absorption, and/or other
optical effect by the a sample on radiation from the excitation
source. For example, the emission may comprise a luminescence or
fluorescence induced by absorption of the excitation
electromagnetic radiation by one or more samples. As used herein
"emission light" refers to an emission comprising an
electromagnetic spectrum having a peak or maximum output (e.g.,
power, energy, or intensity) that is within the visible band of the
electromagnetic spectrum (e.g., electromagnetic radiation within a
wavelength in the range of 420 nanometers to 700 nanometers).
[0024] As used herein, a lens means an optical element configured
to direct or focus incident electromagnetic radiation so as to
converge or diverge such radiation, for example, to provide a real
or virtual image, either at a finite distance or at an optical
infinity. The lens may comprise a single optical element having an
optical power provided by refraction, reflection, and/or
diffraction of the incident electromagnetic radiation.
Alternatively, the lens may comprise a compound system including a
plurality of optical element, for example, including, but not
limited to, an acromatic lens, doublet lens, triplet lens, or
camera lens. The lens may be at least partially housed in or at
least partially enclosed by a lens case or a lens mount.
[0025] As used herein, the term "optical power" means the ability
of a lens or optic to converge or diverge light to provide a focus
(real or virtual) when disposed within air. As used herein the term
"focal length" means the reciprocal of the optical power. As used
herein, the term "diffractive power" or "diffractive optical power"
means the power of a lens or optic, or portion thereof,
attributable to diffraction of incident light into one or more
diffraction orders. Except where noted otherwise, the optical power
of a lens, optic, or optical element is from a reference plane
associated with the lens or optic (e.g., a principal plane of an
optic).
[0026] As used herein, the term "biological sample" means a sample
or solution containing any type of biological chemical or component
and/or any target molecule of interest to a user, manufacturer, or
distributor of the various embodiments of the present invention
described or implied herein, as well as any sample or solution
containing related chemicals or compounds used for the purpose of
conducting a biological assay, experiment, or test. These
biological chemicals, components, or target molecules may include,
but are not limited to, DNA sequences (including cell-free DNA),
RNA sequences, genes, oligonucleotides, molecules, proteins,
biomarkers, cells (e.g., circulating tumor cells), or any other
suitable target biomolecule. A biological sample may comprise one
or more of at least one target nucleic acid sequence, at least one
primer, at least one buffer, at least one nucleotide, at least one
enzyme, at least one detergent, at least one blocking agent, or at
least one dye, marker, and/or probe suitable for detecting a target
or reference nucleic acid sequence. In various embodiments, such
biological components may be used in conjunction with one or more
PCR methods and systems in applications such as fetal diagnostics,
multiplex dPCR, viral detection, and quantification standards,
genotyping, sequencing assays, experiments, or protocols,
sequencing validation, mutation detection, detection of genetically
modified organisms, rare allele detection, and/or copy number
variation.
[0027] According to embodiments of the present invention, one or
more samples or solutions containing at least one biological
targets of interest may be contained in, distributed between, or
divided between a plurality of a small sample volumes or reaction
regions (e.g., volumes or regions of less than or equal to 10
nanoliters, less than or equal to 1 nanoliter, or less than or
equal to 100 picoliters). The reaction regions disclosed herein are
generally illustrated as being contained in wells located in a
substrate material; however, other forms of reaction regions
according to embodiments of the present invention may include
reaction regions located within through-holes or indentations
formed in a substrate, spots of solution distributed on the surface
a substrate, samples or solutions located within test sites or
volumes of a capillary or microfluidic system, or within or on a
plurality of microbeads or microspheres.
[0028] While devices, instruments, systems, and methods according
to embodiments of the present invention are generally directed to
dPCR and qPCR, embodiments of the present invention may be
applicable to any PCR processes, experiment, assays, or protocols
where a large number of reaction regions are processed, observed,
and/or measured. In a dPCR assay or experiment according to
embodiments of the present invention, a dilute solution containing
at least one target polynucleotide or nucleotide sequence is
subdivided into a plurality of reaction regions, such that at least
some of these reaction regions contain either one molecule of the
target nucleotide sequence or none of the target nucleotide
sequence. When the reaction regions are subsequently thermally
cycled in a PCR protocol, procedure, assay, process, or experiment,
the reaction regions containing the one or more molecules of the
target nucleotide sequence are greatly amplified and produce a
positive, detectable detection signal, while those containing none
of the target(s) nucleotide sequence are not amplified and do not
produce a detection signal, or a produce a signal that is below a
predetermined threshold or noise level. Using Poisson statistics,
the number of target nucleotide sequences in an original solution
distributed between the reaction regions may be correlated to the
number of reaction regions producing a positive detection signal.
In some embodiments, the detected signal may be used to determine a
number, or number range, of target molecules contained in the
original solution. For example, a detection system may be
configured to distinguish between reaction regions containing one
target molecule and reaction regions containing two or at least two
target molecules. Additionally or alternatively, the detection
system may be configured to distinguish between reaction regions
containing a number of target molecules that is at or below a
predetermined amount and reaction regions containing more than the
predetermined amount. In certain embodiments, both qPCR and dPCR
processes, assays, or protocols are conducted using a single the
same devices, instruments, or systems, and methods.
[0029] Referring to FIG. 1, a system, apparatus, or instrument 100
for biological analysis comprises one or more of an electronic
processor, computer, or controller 200, a base, mount, or sample
block assembly 300 configured to receive and/or processes a
biological or biochemical sample, and/or an optical system,
apparatus, or instrument 400. Without limiting the scope of the
present invention, system 100 may comprise a sequencing instrument,
a polymerase chain reaction (PCR) instrument (e.g., a real-time PCR
(qPCR) instrument and/or digital PCR (dPCR) instrument), capillary
electrophoresis instrument, an instrument for providing genotyping
information, or the like.
[0030] Electronic processor 200 is configured to control, monitor,
and/or receive data from optical system 400 and/or base 300.
Electronic processor 200 may be physically integrated into optical
system 400 and/or base 300. Additionally or alternatively,
electronic processor 200 may be separate from optical system 400
and base 300, for example, an external desktop computer, laptop
computer, notepad computer, tablet computer, or the like.
Communication between electronic processor 200 and optical system
400 and/or base 300 may be accomplished directly via a physical
connection, such as a USB cable or the like, and/or indirectly via
a wireless or network connection (e.g., via Wi-Fi connection, a
local area network, internet connection, cloud connection, or the
like). Electronic processor 200 may include electronic memory
storage containing instructions, routines, algorithms, test and/or
configuration parameter, test and/or experimental data, or the
like. Electronic processor 200 may be configured, for example, to
operate various components of optical system 400 or to obtain
and/or process data provided by base 300. For example, electronic
processor 200 may be used to obtain and/or process optical data
provided by one or more photodetectors of optical system 400.
[0031] In certain embodiments, electronic processor 200 may
integrated into optical system 400 and/or base 300. Electronic
processor 200 may communicate with external computer and/or
transmit data to an external computer for further processing, for
example, using a hardwire connection, a local area network, an
internet connection, cloud computing system, or the like. The
external computer may be physical computer, such as a desktop
computer, laptop computer, notepad computer, tablet computer, or
the like, that is located in or near system 100. Additionally or
alternatively, either or both the external computer and electronic
processor 200 may comprise a virtual device or system, such as a
cloud computing or storage system. Data may be transferred between
the two via a wireless connection, a cloud storage or computing
system, or the like. Additionally or alternatively, data from
electronic processor 200 (e.g., from optical system 400 and/or base
300) may be transferred to an external memory storage device, for
example, an external hard drive, a USB memory module, a cloud
storage system, or the like.
[0032] In certain embodiments, base 300 is configured to receive a
sample holder or sample carrier 305. Sample holder 305 may comprise
a plurality or array of spatially separated reaction regions,
sites, or locations 308 for containing a corresponding plurality or
array of biological or biochemical samples 310. Reaction regions
308 may comprise any plurality of volumes or locations isolating,
or configured to isolate, the plurality of biological or
biochemical samples 310. For example, reaction regions 308 may
comprise a plurality of through-hole or well in a substrate or
assembly (e.g., sample wells in a standard microtiter plate), a
plurality of sample beads, microbeads, or microspheres in a
channel, capillary, or chamber, a plurality of distinct locations
in a flow cell, a plurality of sample spots on a substrate surface,
or a plurality of wells or openings configured to receive a sample
holder (e.g., the cavities in a sample block assembly configured to
receive a microtiter plate).
[0033] Base 300 may comprise a sample block assembly configured to
control the temperature of sample holder 305 and/or biological
samples 310. Sample block assembly 300 may comprise one or more of
a sample block, a Peltier device or other apparatus for controlling
or cycling temperature, and/or a heat sink (e.g., for aiding in
stabilizing a temperature). Base 300 may comprise a thermal
controller or thermal cycler, for example, to provide or perform a
PCR assay.
[0034] Reaction apparatus 300 may include sample holder 305. At
least some of the reaction regions 308 may include the one or more
biological samples 310. Biological or biochemical samples 310 may
include one or more of at least one target nucleic acid sequence,
at least one primer, at least one buffer, at least one nucleotide,
at least one enzyme, at least one detergent, at least one blocking
agent, or at least one dye, marker, and/or probe suitable for
detecting a target or reference nucleic acid sequence. Sample
holder 305 may be configured to perform at least one of a PCR
assay, a sequencing assay, or a capillary electrophoresis assay, a
blot assay. In certain embodiments, sample holder 305 may comprise
one or more of a microtiter plate, substrate comprising a plurality
of wells or through-holes, a substrate comprising a one or more
channels or capillaries, or a chamber comprising plurality of beads
or spheres containing the one or more biological samples. Reaction
regions 308 may comprise one or more of a plurality of wells, a
plurality of through-holes in substrate, a plurality of distinct
locations on a substrate or within a channel or capillary, a
plurality of microbeads or microspheres within a reaction volume,
or the like. Sample holder 305 may comprise a microtiter plate, for
example, wherein reaction regions 308 may comprise at least 96
well, at least 384, or at least 1536 wells.
[0035] In certain embodiments, sample holder 305 may comprise a
substrate including a first surface, an opposing second surface,
and a plurality of through-holes disposed between the surfaces, the
plurality of through-holes configured to contain the one or more
biological samples, for example as discussed in Patent Application
Publication Numbers US 2014-0242596 and WO 2013/138706, which
applications are herein incorporated by reference as if fully set
forth herein. In such embodiments, the substrate may comprise at
least 3096 through-holes or at least 20,000 through-holes. In
certain embodiments, sample holder 305 may comprise an array of
capillaries configured to pass one or more target molecules or
sequence of molecules.
[0036] In certain embodiments, system 100 may optionally include a
heated or temperature controlled cover 102 that may be disposed
above sample holder 305 and/or base 300. Heated cover 102 may be
used, for example, to prevent condensation above the samples
contained in sample holder 305, which can help to maintain optical
access to biological samples 310.
[0037] In certain embodiments, optical system 400 comprises an
excitation source, illumination source, radiation source, or light
source 402 that produces at least a first excitation beam 405a
characterized by a first wavelength and a second excitation beam
405b characterized by a second wavelength that is different from
the first wavelength. Optical system 400 also comprises an optical
sensor or optical detector 408 configured to receive emissions or
radiation from one or more biological samples in response to
excitation source 410 and/or to one or more of excitation beams
405a, 405b. Optical system 400 additionally comprises an excitation
optical system 410 disposed along an excitation optical path 412
between excitation source 402 and one or more biological samples to
be illuminated. Optical system 400 further comprises an emission
optical system 415 disposed along an emission optical path 417
between the illuminated sample(s) and optical sensor 408. In
certain embodiments, optical system 400 may comprise a beamsplitter
420. Optical system 400 may optionally include a beam dump or
radiation baffle 422 configured reduce or prevent reflection of
radiation into emission optical path 417 from excitation source 402
that impinges on beamsplitter 420.
[0038] In the illustrated embodiment shown in FIG. 1, as well as
other embodiments of the invention disclosed herein, excitation
source 402 comprises a radiation source 425. Radiation source 425
may comprise one or more of at least one an incandescent lamp, at
least one gas discharge lamp, at least one light emitting diode
(LED), at least one organic light emitting diode, and/or at least
one laser. For example, radiation source 425 may comprise at least
one Halogen lamp, Xenon lamp, Argon lamp, Krypton lamp, diode
laser, Argon laser, Xenon laser, excimer laser, solid-state laser,
Helium-Neon laser, dye laser, or combinations thereof. Radiation
source 425 may comprise a light source characterized by a maximum
or central wavelength in the visible band of the electromagnetic
spectrum. Additionally or alternatively, radiation source 425 may
comprise an ultraviolet, infrared, or near-infrared source with a
corresponding maximum or central wavelength within on one of those
wavelength bands of the electromagnetic spectrum. Radiation source
425 may be a broadband source, for example, having a spectral
bandwidth of at least 100 nanometers, at least 200 nanometers, or
at least 300 nanometers, where the bandwidth is defined as a range
over which the intensity, energy, or power output is greater than a
predetermined amount (e.g., where the predetermined amount is at or
about 1%, 5%, or 10% of a maximum or central wavelength of the
radiation source). Excitation source 402 may additionally comprise
a source lens 428 configured to condition emissions from radiation
source 425, for example, to increase the amount of excitation
radiation received at sample holder 305 and/or into biological
samples 310. Source lens 428 may comprise a simple lens or may be a
compound lens including two or more elements.
[0039] In certain embodiments, excitation source 402 further
comprises two or more excitation filters 430 moveable into and out
of excitation optical path 412, for instance, used in combination
with a broadband excitation source 402. In such embodiments,
different excitation filters 430 may be used to select different
wavelength ranges or excitation channels suitable for inducing
fluorescence from a respective dye or marker within biological
samples 310. One or more of excitation filters 430 may have a
wavelength bandwidth that is at least .+-.10 nanometers or at least
.+-.15 nanometers. Excitation filters 430 may comprise a plurality
of filters that together provide a plurality of band passes
suitable for fluorescing one or more of a SYBR.RTM. dye or probe, a
FAM.TM. dye or probe, a VIC.RTM. dye or probe, a ROX.TM. dye or
probe, or a TAMRA.TM. dye or probe. Excitation filters 430 may be
arrange in a rotatable filter wheel (not shown) or other suitable
device or apparatus providing different excitation channels using
excitation source 402. In certain embodiments, excitation filters
430 comprise at least 5 filter or at least 6 filter.
[0040] In certain embodiments, excitation source 402 may comprise a
plurality of individual excitation sources that may be combined
using one more beamsplitters or beam combiners, such that radiation
from each individual excitation source is transmitted along a
common optical path, for example, along excitation optical path 412
shown in FIG. 1. Alternatively, at least some of the individual
excitation sources may be arranged to provided excitation beams
that propagate along different, non-overlapping optical paths, for
example, to illuminate different reaction regions of the plurality
of reaction regions 308. Each of the individual excitation sources
may be addressed, activated, or selected to illuminate reaction
regions 308, for example, either individually or in groups or all
simultaneously. In certain embodiments, the individual excitation
sources may be arrange in a one-dimensional or two-dimensional
array, where one or more of the individual excitation sources is
characterized by a maximum or central wavelength that is different
than that of at least one of the other individual excitation
sources in the array.
[0041] In certain embodiments, first excitation beam 405a comprises
a first wavelength range over which an intensity, power, or energy
of first excitation beam 405a is above a first predetermined value
and second excitation beam 405b comprises a second wavelength range
over which an intensity, power, or energy of second excitation beam
405b is above a second predetermined value. The characteristic
wavelength of the excitation beams 405a, 405b may be a central
wavelength of the corresponding wavelength range or a wavelength of
maximum electromagnetic intensity, power, or energy over the
corresponding wavelength range. The central wavelengths of at least
one of the excitation beams 405 may be an average wavelength over
the corresponding wavelength range. For each excitation beam 405
(e.g., excitation beams 405a, 405b), the predetermined value may be
less than 20% of the corresponding maximum intensity, power, or
energy; less than 10% of the corresponding maximum intensity,
power, or energy; less than 5% of the corresponding maximum
intensity, power, or energy; or less than 1% of the corresponding
maximum intensity, power, or energy. The predetermined values may
be the same for all excitation beams 405 (e.g., for both excitation
beams 405a, 405b) or the predetermined values may be different from
one another. In certain embodiments, the wavelength ranges of the
first and second excitation beams 405a, 405b do not overlap, while
in other embodiments at least one of the wavelength ranges at least
partially overlaps that of the other. In certain embodiments, the
first and second central wavelengths are separated by at least 20
nanometer. In certain embodiments, at least one of the first and
second wavelength ranges has a value of at least 20 nanometer or at
least 30 nanometers.
[0042] Excitation optical system 410 is configured to direct
excitation beams 405a, 405b to the one or more biological samples.
Where applicable, references herein to excitation beams 405a, 405b
may be applied to embodiment comprising more than two excitation
beams 405. For example, excitation source 402 may be configured to
direct at least five or six excitation beams 405. Excitation beams
405a, 405b may be produced or provided simultaneously, may be
temporally separated, and/or may be spatially separated (e.g.,
wherein excitation beams 405a is directed to one reaction region
308 and excitation beams 405b is directed to a different reaction
region 308). The excitation beams 405 may be produced sequentially,
for example, by sequentially turning on and off different-colored
individual radiation source 425 that are characterized by different
wavelengths or by sequentially placing different color filters in
front of a single radiation source 425. Alternatively, excitation
beams 405a, 405b may be produced simultaneously, for example, by
using a multi-wavelength band filter, beamsplitter, or mirror, or
by coupling together different individual radiation source 425,
such as two different-colored light emitting diodes (LEDs). In some
embodiments, excitation source 402 produces more than two
excitation beams 405, wherein excitation optical system 410 directs
each of the excitation beams to one or more biological samples 310.
Referring to FIG. 2, excitation source 402 may comprise at least 5
individual radiant sources 425a, 425b, 425c, 425d, 425e that are
combined to transmit along a common excitation optical path 412.
Excitation source 402 may also comprise corresponding sources
lenses 428a, 428b, 428c, 428d, 428e. Radiation from radiant sources
425a, 425b, 425c, 425d, 425e may be combined using a plurality of
combiner optical elements 429a, 429b, 429c. Combiner optical
elements 429a, 429b, 429c may comprise one or more of a neutral
density filter, a 50/50 beamsplitter, a dichroic filter or mirror,
a cube beamsplitter, or the like. Combiner optical elements 429a,
429b, 429c are one example of how to combine various individual
sources 425 and it will be appreciated that other combinations and
geometrical arrangements of individual radiant sources 425 and
combiner optical elements 429 are within the scope of embodiments
of the present invention. One or more of individual radiant sources
425a, 425b, 425c, 425d, 425e may be characterized by a central
wavelength and/or wavelength range that is differ from that of the
other individual radiant sources 425a, 425b, 425c, 425d, 425e.
[0043] Referring to FIGS. 3-4, the spectral distribution of
radiation source 425 may be selected in a non-obvious manner to
enable at least five excitation beams 405 of different colors or
excitation channels to be used with one common beamsplitter 420,
while simultaneously maintaining acceptable or predetermined data
throughput for all excitation channels, for example, during each
cycle of the qPCR assay. As used herein, the term "excitation
channel" means each of several, distinct electromagnetic wavelength
bands providing by an excitation source (e.g., excitation source
402) that are configured to illuminate one or more biological
samples (e.g., biological samples 310). As used herein, the term
"emission channel" means each of several, distinct emission
wavelength bands over which electromagnetic radiation is allowed to
pass onto an optical sensor or detector (e.g., optical sensor
408).
[0044] FIG. 3 shows the relative energy over the wavelength
spectrum for three different radiation sources. The dashed line
plot is the spectrum of a Halogen lamp (herein referred to as
"Source 1") characterized by relatively low energy levels in the
blue wavelength range of the visible spectrum and increasing energy
until a peak at about 670 nanometers. The dash-dot spectrum plot is
that of a commercially available LED light source (herein referred
to as "Source 2"), which has peak energy at around 450 nanometers
and a lower peak from about 530 nanometers to about 580 nanometers,
then steadily decreasing energy into the red wavelength range of
the visible spectrum. The solid line plot is the spectrum of
another LED light source (herein referred to as "Source 3")
according to an embodiment of the present invention (e.g., an
exemplary spectrum for excitation source 402). FIG. 4 shows
integrated energy over various excitation channels for each of the
three sources shown in FIG. 3, where the spectrums for these
channels are those of typical excitation filter used in the field
of qPCR. The wavelength ranges and excitation filter designations
are shown below in Table 1, where X1 is excitation channel 1, X2 is
excitation channel 2, and so forth.
TABLE-US-00001 TABLE 1 Spectral bandwidth of excitation filters
used in FIG. 4. Excitation Wavelength Filter Range Channel
(nanometers) X1 455-485 X2 510-530 X3 540-560 X4 570.5-589.5 X5
630.5-649.5 X6 650-674
[0045] In the field of qPCR, one important performance parameter is
the total time to obtain emission data for samples containing
multiple target dyes. For example, in some cases it is desirable to
obtain emission data from multiple dyes or probes over one or more
emission channels, designated M1-M6, for each excitation channel
used to illuminate the sample(s) (e.g., M1-M6 with X1, M2-M6 with
X2, M3-M6 with X3, M4-M6 with X4, M5-M6 with X5, and/or M6 with
X6). The inventors have found that when Source 2 is used in a
system having a single, broadband beamsplitter for five or six
excitation/emission filter channels (e.g., excitation channels
X1-X6 with combinations emission channels M1-M6), the amount of
time to obtain data for excitation channel 5 and/or excitation
channel 6 could be unacceptably long for certain applications. To
remedy this situation, it is possible to use one or more narrow
band, dichroic beamsplitters for excitation channels 1 and/or 2 to
increase the amount of excitation light receive by the sample(s),
and the amount of emission light received by the sensor (so that
the overall optical efficiency is increased by using dichroic beam
splitter, in this case). However, this precludes the use of a
single beamsplitter arrangement, as shown in FIG. 1 and, therefore,
the corresponding advantages of a single beamsplitter configuration
(e.g., reduced size, cost, complexity) are lost. A better solution
has been discovered in which a light source, such as Source 3, is
used in combination with a single beamsplitter (e.g., a broadband
beamsplitter such as a 50/50 beamsplitter), such as beamsplitter
420. It has been found that the relative energy in excitation
channels X1, X5, and/or X6 may be used to identify an excitation
source 402 suitable for use with a single beamsplitter embodiment
to provide acceptable total integration time for collecting
emission data over five or six excitation channels. Using LED
Source 2 and LED Source 3 as examples, the following data shown in
Table 2 below may be derived for the data shown in FIGS. 3 and
4.
TABLE-US-00002 TABLE 2 Normalized LED intensity of each filter
channel with normalization over channel 2. Ratio Source 2 Source 3
X1/X2 2.02 3.00 X2/X2 1.00 1.00 X3/X2 1.20 0.98 X4/X2 1.09 0.89
X5/X2 0.49 0.90 X6/X2 0.38 0.90
[0046] Based on such data, the inventors have found that, in
certain embodiments, improved performance (e.g., in terms of
shorter Channel 1 integration time) may be obtain when X1/X2 is
greater than 2.02 (e.g., greater than or equal to 3). Additionally
or alternatively, in other embodiments, improved performance (e.g.,
in terms of shorter Channel 1 integration time) may be obtain when
X5/X2 is greater than 0.49 (for example, greater than or equal to
0.9) and/or when X6/X2 is greater than 0.38 (for example, greater
than or equal to 0.9). For the criteria set forth here, "X1" means
an excitation channel that has a spectral output characterized by a
maximum power, energy, or intensity within the wavelength band
including 455-485 nanometers; "X2" means an excitation channel that
has a spectral output characterized by a maximum power, energy, or
intensity within the wavelength band including 510-530 nanometers;
"X5" means an excitation channel that has a spectral output
characterized by a maximum power, energy, or intensity within the
wavelength band including 630.5-649.5 nanometers; "X6" means an
excitation channel that has a spectral output characterized by a
maximum power, energy, or intensity within the wavelength band
including 650-674 nanometers
[0047] Referring again to FIG. 1, excitation beams 405 are directed
along excitation optical path 412 during operation toward sample
processing base 300, for example, toward reaction regions 308 when
sample holder 305 is present. When present, source lens 428 is
configure to condition excitation beams 405, for example, to
capture and direct a large portion of the emitted radiation from
excitation source 402. In certain embodiments, one or more mirrors
432 (e.g., fold mirrors) may be incorporated along excitation
optical path 412, for example, to make optical system 400 more
compact and/or to provide predetermined package dimensions. FIG. 1
illustrated one mirror 432; however, addition mirrors may be used,
for example to meet packaging design constraints. As discussed in
greater detail below herein, additional lenses may be disposed near
sample holder 305, for example, in order to further condition the
excitation beams 405 and/or corresponding emissions from biological
samples contained in one or more reaction regions.
[0048] Emission optical system 415 is configured to direct
emissions from the one or more biological samples to optical sensor
408. At least some of the emissions may comprise a fluorescent
emission from at least some of the biological samples in response
to at least one of the excitation beams 405. Additionally or
alternatively, at least some of the emissions comprise radiation
from at least one of the excitation beams 405 that is reflected,
refracted, diffracted, scattered, or polarized by at least some of
the biological samples. In certain embodiments, emission optical
system 415 comprise one or more emission filters 435 configured,
for example, to block excitation radiation reflected or scattered
into emission optical path 417. In certain embodiments, there is a
corresponding emission filter 435 for each excitation filter 430.
Referring to FIG. 8, in certain embodiments, the excitation filter
430 are arranged in an excitation filter wheel 431 and/or the
emission filters 435 are arranged in an emission filter wheel
436.
[0049] In certain embodiments, emission optical system 415
comprises a sensor lens 438 configured to direct emissions from at
least some of the biological samples onto optical sensor 408.
Optical sensor 408 may comprise a single sensor element, for
example, a photodiode detector or a photomultiplier tube, or the
like. Additionally or alternatively, optical sensor 408 may
comprise an array sensor including an array of sensors or pixels.
Array sensor 408 may comprise one or more of a complementary
metal-oxide-semiconductor sensor (CMOS), a charge-coupled device
(CCD) sensor, a plurality of photodiodes detectors, a plurality of
photomultiplier tubes, or the like. Sensor lens 438 may be
configured to from an image from the emissions from one or more of
the plurality of biological samples 310. In certain embodiments,
optical sensor 408 comprises two or more array sensors 408, for
example, where two or more images are formed from the emissions
from one or more of the plurality of biological samples 310. In
such embodiments, emissions from one or more of the plurality of
biological samples 310 may be split to provide two signals of the
one or more of the plurality of biological samples 310. In certain
embodiments, the optical sensor comprises at least two array
sensors.
[0050] Beamsplitter 420 is disposed along both excitation and
emission optical paths 412, 417 and is configured to receive both
first and second excitation beams 405a, 405b during operation. In
the illustrated embodiment shown in FIG. 1, beamsplitter 420 is
configured to transmit the excitation beams 405 and to reflect
emissions from the biological samples 310. Alternatively,
beamsplitter 420 may be configured to reflect the excitation beams
and to transmit emissions from the biological samples 310. In
certain embodiments, beamsplitter 420 comprises a broadband
beamsplitter having the same, or approximately the same,
reflectance for all or most of the excitation beams 405 provided by
excitation source 402 and directed to the reaction regions 308
(e.g., excitation beams 405a, 405b in the illustrated embodiment).
For example, beamsplitter 420 may be a broadband beamsplitter
characterized by a reflectance that is constant, or about constant,
over a wavelength band of at least 100 nanometers, over a
wavelength band of at least 200 nanometers, or over the visible
wavelength band of the electromagnetic spectrum, over the visible
and near IR wavelength bands of the electromagnetic spectrum, or
over a wavelength band from 450 nanometers to 680 nanometers. In
certain embodiments, beamsplitter 420 is a neutral density filter,
for example, a filter having a reflectance of, or about, 20%, 50%,
or 80% over visible wavelength band of the electromagnetic
spectrum. In certain embodiments, beamsplitter 420 is a dichroic
beamsplitter that is transmissive or reflective over one or more
selected wavelength ranges, for example, a multi-wavelength band
beamsplitter that is transmissive and/or reflective over more than
one band of wavelengths centers at or near a peak wavelength of
excitation beams 405.
[0051] In certain embodiments, beamsplitter 420 is a single
beamsplitter configure to receive some or all of the plurality of
excitation beams 405 (e.g., excitation beams 405a, 405b), either
alone or in combination with a single beam dump 422. Each
excitation beam may be referred to as an excitation channel, which
may be used alone or in combination to excite different fluorescent
dyes or probe molecule in one or more of the biological samples
310. By contrast many prior art systems and instruments, for
example, in the field of qPCR, provide a plurality of excitation
beams by using a separate beamsplitter and/or beam dump for each
excitation channel and/or each emission channel of the system or
instrument. In such prior art systems and instruments,
chromatically selective dichroic filters are typically used in at
least some of the excitation channels to increase the amount of
radiation received at the samples. Disadvantages of systems and
instruments using different beamsplitters and/or beam dumps for
each channel include an increase in size, cost, complexity, and
response time (e.g., dues to increased mass that must be moved or
rotated when changing between excitation and/or emission channels).
The inventors have discovered that it is possible to replace these
plural beamsplitters and/or beam dumps with the single beamsplitter
420 and/or single beam dump 422, while still providing an
acceptable or predetermined system or instrument performance, for
example, by proper selection of spectral distribution of excitation
source 402 and/or by configuring the systems or instruments to
reduce the amount of stray or unwanted radiation received by
optical sensor 408 (as discuss further herein). Thus, embodiments
of the present invention may be used to provide systems and
instruments that have reduced size, cost, complexity, and response
time as compared to prior art systems and instruments.
[0052] Referring to FIGS. 5 and 6, in certain embodiments, system
100 comprises an instrument housing 105 and sample holder drawer
110 comprising base 300 and configured during use to receive, hold,
or contain sample holder 305 and to position sample holder 305 to
provide optical coupling thereof with optical system 400. With
drawer 110 closed (FIG. 6), housing 105 may be configured to
contain or enclose sample processing system 300 and optical system
400. In certain embodiments, housing 105 may contain or enclose all
or portions of electronic processor 200.
[0053] Referring to FIGS. 7-9, in certain embodiments, optical
system 400 may further comprise a lens 440 and/or a lens array 442,
which may comprise a plurality of lenses corresponding to each of
the reaction regions 308 of sample holder 305. Lens 440 may
comprises a field lens, which may be configured to provide a
telecentric optical system for a least one of sample holder 305,
reaction regions 308, lens array 442, or optical sensor 408. As
shown in illustrated embodiment in FIGS. 7 and 9, lens 440 may
comprise a Fresnel lens.
[0054] Referring again to FIGS. 7 and 9, in certain embodiments,
base 300 comprises a sample block assembly 300 comprising a sample
block 302, temperature controller 303, such as a
[0055] Peltier device 303, and a heat sink 304. Sample block
assembly 300 may be configured to provide a thermal controller or
thermal cycling (e.g., provide a PCR assay or temperature profile),
maintain a temperature of sample holder 305 or biological sample(s)
310, and/or otherwise maintain, control, adjust, or cycle heat flow
or temperature of sample holder 305 or biological sample(s)
310.
[0056] With additional reference to FIGS. 10-14, in certain
embodiments, optical system 400 includes an imaging unit 445
comprising an optical sensor circuit board 448, sensor lens 438
(which may be a compound lens, as illustrated in FIG. 10), an inner
lens mount 449, an outer lens mount 450, a threaded housing 452,
and a focusing gear 455. Optical sensor circuit board 448, threaded
housing 452, and sensor lens 438 together may form a cavity 458
that encloses or contains optical sensor 408 and may be configured
to block any external light from impinging optical sensor 408 that
does not enter through sensor lens 438. Outer lens mount 450
comprises an outer surface containing gear teeth 460 that may be
moveably or slideably engaged with the teeth of focusing gear 455
via a resilient element (not shown), such as a spring. In certain
embodiments, focusing gear 455 moves or slide along a slot 462 of a
plate 465, as illustrated in FIG. 14. Inner lens mount 449
comprises a threaded portion 468 that engages or mates with a
threaded portion of threaded housing 452.
[0057] Inner lens mount 449 may be fixedly mounted to outer lens
mount 450, while threaded housing 452 is fixedly mounted relative
to optical sensor circuit board 448. Inner lens mount 449 is
moveably or rotatably mounted to threaded housing 452. Thus,
focusing gear 455 and outer lens mount 450 may be engaged such that
a rotation of focusing gear 455 also rotates outer lens mount 450.
This, in turn, causes inner lens mount 449 and sensor lens 438 to
move along an optical axis of sensor lens 438 via the threads in
inner lens mount 449 and threaded housing 452. In this manner, the
focus of sensor lens 438 may be adjusted without directly engaging
sensor lens 438 or its associated mounts 449, 450, which are buried
within a very compact optical system 400. Engagement with focusing
gear 455 may be either by hand or automated, for example using a
motor (not shown), such as a stepper motor or DC motor.
[0058] Referring to FIGS. 11 and 13-17, in certain embodiments,
imaging unit 445 further comprises a locking device or mechanism
470. Locking device 470 comprises an edge or tooth 472 that may be
slideably engaged between two teeth of focusing gear 455 (see FIGS.
15-17). As illustrated in FIGS. 15 and 16, locking device 470 may
have a first position (FIG. 15) in which focusing gear 455 is free
to rotate and adjust the focus of sensor lens 438 and a second
position (FIG. 14) is which focusing gear 455 is locked in position
and impeded or prevented from rotating. In this manner, the focus
of sensor lens 438 may be locked while advantageously avoiding
direct locking contact or engagement with threads 468 of inner lens
mount 449, which could damage the threads and prevent subsequent
refocusing of sensor lens 438 after being locked into position.
Operation of locking device 470 may be either manually or in an
automated manner. In certain embodiments, locking mechanism 470
further comprises a resilient element such as a spring (not shown),
wherein rotation of focusing gear 455 may be accomplished by
overcoming a threshold force produced by the resilient element.
[0059] Referring to FIG. 18, optical system 400 may also include an
optics housing 477. In certain embodiments, optical system 400
includes a radiation shield 475 comprising a sensor aperture 478
disposed along emission optical path 417 and at least one blocking
structure 480 disposed to cooperate with sensor aperture 478 such
that the only radiation from excitation beams 405, and reflected
off an illuminated surface or area 482, to pass through sensor
aperture 478 is radiation that has also reflected off at least one
other surface of, or within, the optics housing 477. In other
words, radiation shield 475 is configured such that radiation from
excitation beams 405 reflected illuminated area 482 are blocked
from directly passing through aperture 478 and, therefore, from
passing into sensor lens 438 and onto optical detector 408. In
certain embodiments, illuminated area 482 comprises the area
defined by all the apertures 483 of heated cover 102 corresponding
to the plurality of reaction regions 308.
[0060] In the illustrated embodiment of FIG. 18, blocking structure
480 comprises a shelf 480. Dashed lines or rays 484a and 484b may
be used to illustrate the effectiveness of blocking structure 480
in preventing light directly reflected from illuminated area 482
from passing through sensor aperture 478 and onto senor lens 438
and/or optical sensor 408. Ray 484a originates from an edge of
illuminated area 482 an just passes shelf 480, but does not pass
through sensor aperture 478. Ray 484b is another ray originating
from the same edge of illuminated area 482 that is blocked by shelf
480. As can be seen, this ray would have entered through sensor
aperture 478 were it not for the presences of shelf 480.
[0061] With continued reference to FIG. 18, in certain embodiments,
optical system 400 may further comprise an energy or power
detection unit comprising a power or energy sensors 490 optically
coupled to one end of a light pipe 492. An opposite end 493 of
light pipe 492 is configured to be illuminated by excitation beams
405. Light pipe end 493 may be illuminated either directly by
radiation contained in excitation beams 405 or indirectly, for
example, by radiation scattered by a diffuse surface. In certain
embodiments, sensor 490 is located outside of the excitation
optical path 412 from excitation source 402. Additionally or
alternatively, sensor 490 is located outside optics housing 477
and/or is located at a remote location outside instrument housing
105. In the illustrated embodiment shown in FIG. 18, light pipe end
493 is disposed near or adjacent mirror 432 and may be oriented so
that the face of the light pipe is perpendicular, or nearly
perpendicular, to the surface of mirror 432 that reflects
excitation beams 405. The inventors have discovered that the low
amount of energy or power intercepted by light pipe 492 when
oriented in this way is sufficient for the purpose of monitoring
the energy or power of excitation beams 405. Advantageously, by
locating sensor 490 outside the optical path of excitation beams a
more compact optical system 400 may be provided.
[0062] In certain embodiments, light pipe 492 comprises a single
fiber or a fiber bundle. Additionally or alternatively, light 492
may comprise a rod made of a transparent or transmissive material
such as glass, Plexiglas, polymer based material such as acrylic,
or the like.
[0063] Referring to FIGS. 19 and 20, in certain embodiments
instrument 100 comprises a position source 500 configured to emit
radiation 502 and a corresponding position sensor 505 configured to
receive radiation 502 from position source 500. Position source 500
and position sensor 505 may be configured to produce a position
signal indicative of a position of an optical element 435 disposed
along an optical paths. In certain embodiments, instrument 100 may
further comprise a radiation shield 510 configured to block at
least some radiation 502 from position source 505.
[0064] The above presents a description of the best mode
contemplated of carrying out the present invention, and of the
manner and process of making and using it, in such full, clear,
concise, and exact terms as to enable any person skilled in the art
to which it pertains to make and use this invention. This invention
is, however, susceptible to modifications and alternate
constructions from that discussed above which are fully equivalent.
Consequently, it is not the intention to limit this invention to
the particular embodiments disclosed. On the contrary, the
intention is to cover modifications and alternate constructions
coming within the spirit and scope of the invention as generally
expressed by the following claims, which particularly point out and
distinctly claim the subject matter of the invention.
[0065] Exemplary systems for methods related to the various
embodiments described in this document include those described in
following applications: [0066] U.S. design patent application Ser.
No. 29/516,847, filed on Feb. 6, 2015; and [0067] U.S. design
patent application Ser. No. 29/516,883; filed on Feb. 6, 2015; and
[0068] U.S. provisional patent application No. 62/112,910, filed on
Feb. 6, 2015; and [0069] U.S. provisional patent application No.
62/113,006, filed on Feb. 6, 2015; and [0070] U.S. provisional
patent application No. 62/113,183, filed on Feb. 6, 2015; and
[0071] U.S. provisional patent application No. 62/113,077, filed on
Feb. 6, 2015; and [0072] U.S. provisional patent application No.
62/113,058, filed on Feb. 6, 2015; and [0073] U.S. provisional
patent application No. 62/112,964, filed on Feb. 6, 2015; and
[0074] U.S. provisional patent application No. 62/113,118, filed on
Feb. 6, 2015; and [0075] U.S. provisional patent application No.
62/113,212, filed on Feb. 6, 2015; and [0076] U.S. patent
application Ser. No. ______ (Life Technologies Docket Number
LT01023), filed on Feb. 5, 2016; and [0077] U.S. patent application
Ser. No. ______ (Life Technologies Docket Number LT01024), filed on
Feb. 5, 2016; and [0078] U.S. patent application Ser. No. ______
(Life Technologies Docket Number LT01025), filed on Feb. 5, 2016;
and [0079] U.S. patent application Ser. No. ______ (Life
Technologies Docket Number LT01028), filed on Feb. 5, 2016; and
[0080] U.S. patent application Ser. No. ______ (Life Technologies
Docket Number LT01029), filed on Feb. 5, 2016; and [0081] U.S.
patent application Ser. No. ______ (Life Technologies Docket Number
LT01032), filed on Feb. 5, 2016; and [0082] U.S. patent application
Ser. No. ______ (Life Technologies Docket Number LT01033), filed on
Feb. 5, 2016, all of which are also herein incorporated by
reference in their entirety.
* * * * *