U.S. patent number 3,607,659 [Application Number 04/786,570] was granted by the patent office on 1971-09-21 for fermentation apparatus.
This patent grant is currently assigned to Temple University of the Commonwealth System of Higher Education. Invention is credited to James L. Bloomer.
United States Patent |
3,607,659 |
Bloomer |
September 21, 1971 |
FERMENTATION APPARATUS
Abstract
Apparatus is provided in the form of fermentation vessels which
are provided with caps for substantially sealing the same against
contamination, and with tube means for communicating between the
inside and outside of the vessels. Stirring is effected by a
magnetic element disposed within the vessels which is actuated by
an external electromagnetic source.
Inventors: |
Bloomer; James L.
(Philadelphia, PA) |
Assignee: |
Temple University of the
Commonwealth System of Higher Education (Philadelphia,
PA)
|
Family
ID: |
25138964 |
Appl.
No.: |
04/786,570 |
Filed: |
December 24, 1968 |
Current U.S.
Class: |
435/302.1 |
Current CPC
Class: |
C12M
23/08 (20130101); C12M 27/02 (20130101); C12M
23/38 (20130101); C12M 37/02 (20130101) |
Current International
Class: |
C12M
1/24 (20060101); C12b 001/00 () |
Field of
Search: |
;195/139,143,142,127,139LE ;23/292 ;215/56 |
References Cited
[Referenced By]
U.S. Patent Documents
Foreign Patent Documents
Primary Examiner: Tanenholtz; Alvin E.
Claims
I claim:
1. An apparatus for use in organism growth comprising an
open-topped vessel having a body portion and having a neck portion
of reduced diameter, an open ended cap having side and end walls
and being disposed in inverted relation over said vessel open end,
with lower sidewall portions of said cap being in sufficiently
close fitted relation to an adjacent outer portion of said vessel
that gaseous outflow therebetween from inside the vessel will
maintain sterility inside the vessel, including tube means for
providing gas inflow to the interior of the vessel; said tube means
comprising a duct extending from outside the vessel, passing
between said close fitted cap and vessel portions, extending
between said cap and vessel to the upper end of said vessel, being
reversely bent beneath the cap endwall and over the vessel open
upper end, and extending downwardly into a lower interior portion
of said vessel, with at least one of an innoculation and sampling
tube means also being provided and comprising an additional duct
extending from outside the vessel, passing from outside said cap
between said close fitted cap and vessel portions, extending
between said cap and vessel to the upper end of said vessel, being
reversely bent beneath the cap endwall andover the vessel open
upper end, and extending downwardly into a lower interior portion
of said vessel, wherein said gas inflow tube means includes gas
filter means and means for connection to an external gas source,
and wherein removable cover means for each said innoculation and
sampling tube means is provided.
2. The apparatus of claim 1, including a magnetic stirring element
disposed within the vessel, and adapted for use with an external
magnetic actuator for rotating the same.
3. The apparatus of claim 1, wherein a diffusing frit is provided
on the lowermost end of said inflow tube means inside said
vessel.
4. The apparatus of claim 1, wherein said additional duct has a
second reverse bend around the lower end of the inverted cap and
terminates in an upwardly extending free end.
5. The apparatus of claim 1, wherein both said innoculation and
sampling tube means are provided.
6. The apparatus of claim 1, wherein each said tube means is
angularly outwardly bent inside said vessel near the lower end
thereof.
7. An apparatus for use in organism growth, comprising an
open-topped vessel having body portion and having an open upper end
of reduced diameter relative to the vessel bottom, with the vessel
body diameter progressing to a greatest diameter at the bottom
thereof, and with a cap being provided having an open lower end,
and being disposed over the vessel open upper end in inverted
close-fitting relation thereto, with the cap sidewalls extending
downwardly along, but outside the upper ends of the vessel
sidewalls, and with the cap upper end being closed except for the
passage of a gas inlet tube means therethrough in heat sealed
relation thereto, with the gas inlet tube means comprising a duct
extending through the upper end wall of the cap, and down into the
lower interior portion of the vessel from outside the cap,
comprising means for providing gas to the interior of said vessel,
and with at least one of an innoculation and sampling tube means
permanently connected to the vessel at the bottom thereof, in
integrally sealed relation therewith, and extending vertically
upwardly therefrom, and terminating in an upwardly opening free end
having a removable cover therefor, to comprise means for providing
access to the interior bottom portion of said vessel, and wherein
said gas inflow tube means includes means for connection to an
external gas source, and wherein filtering means are provided
within said gas inlet tube means.
8. The apparatus of claim 7, wherein a magnetic stirring element is
disposed within the vessel, and is adapted for use with an external
device adapted for rotating the magnetic stirring element from
outside the vessel.
9. The device of claim 7, wherein both said innoculation and
sampling tube means are provided.
10. The device of claim 7, wherein said one of said innoculation
and sampling tube means is provided with removable filtration means
disposed therein.
11. The device of claim 7, wherein said gas inlet tube means is
provided with a diffusing frit at its lowermost portion inside the
vessel, for distribution of gas passing through the tube means into
a liquid contained within the vessel.
12. The apparatus of claim 7, wherein said gas inlet tube means is
angularly bent in an outward direction, at the lowermost interior
end thereof, within the vessel.
Description
BACKGROUND OF THE INVENTION
In fermentation vessels (and other vessels used for organism
growth) of the prior art, several apparatus have been developed,
principally for use in analyzing organic mixtures while the
mixtures are in motion, as a continuous process. Generally, such
apparatus have been rather complicated, and have required the use
of rotary seals, principally for providing stirring from an
external source, such seals often sticking when the vessels are
used for many solutions, such as sugar solutions.
Furthermore, there has developed a need for apparatus which is
sufficiently inexpensive to be used in quantity, as for example,
for laboratory use by students and the like, but which will provide
sufficient safeguards against contamination when used for such
studies as culture growths, fungal metabolic studies and the like.
In such studies, it is often desirable to inoculate the culture or
growth, and it is additionally desirable to often take samples of
the growth at various stages of its development. A primary
consideration at all times is that contamination of the apparatus
must be prevented, and in this regard, along with the other
above-discussed parameters, it has been necessary to develop
vessels which would satisfy all of the requirements mentioned
above, but which would still be economical, as well as readily
capable of being assembled.
SUMMARY OF THE INVENTION
The present invention seeks to provide an apparatus which may be
economically manufactured and assembled, to provide
contamination-free apparatus for various studies, wherein
inoculation and sampling facilities are provided for the apparatus,
by using a vessel and a cap fitted over its upper end, sufficiently
snug with the top of the vessel, but wherein means are provided for
delivering a gas, such as sterile air to the vessel, the outflow of
such gas from the vessel providing a seal against contamination
from the ambient air. Novel means are also provided whereby gas
inlet, inoculation and sampling tubes communicate between the
inside and outside of the vessel.
Accordingly, it is a primary object of this invention to provide a
novel fermentation apparatus, whereby a gas seal is provided
between a vessel and its cap, but whereby the apparatus remains
free from contamination during use, and also whereby access is
provided between the interior of the vessel and externally
thereof.
It is another object of this invention to provide a novel apparatus
comprising a vessel and a cap therefore, whereby tubes extend from
the inside of the vessel to the outside thereof, such tubes being
closely fitted with the vessel cap for purposes of preventing
contamination of the vessel.
It is a further object of this invention to provide a novel vessel
having a cap, wherein the vessel is provided with inoculation,
sampling and gas inlet tubes, as separate communication routes
between the inside and the outside of the vessel.
It is a further object of this invention to accomplish the above
object, wherein magnetic stirring means are utilized interiorly of
the vessel.
Other objects and advantages of the present invention will become
readily apparent to those skilled in the art from a reading of the
following brief description of the drawing figures, detailed
description of the preferred embodiments and the appended
claims.
IN THE DRAWINGS
FIG. 1 is a vertical sectional view, taken through the apparatus of
this invention, generally along the line I--I of FIG. 2.
FIG. 2 is a transverse sectional view taken through the apparatus
of FIG. 1, generally along the line II--II, wherein several of the
components of the apparatus are illustrated in plan view.
FIG. 3 is a vertical sectional view of an alternative form of the
apparatus of this invention.
FIG. 4 is a transverse sectional view of the apparatus of FIG. 3,
taken generally along the lines IV--IV of FIG. 3.
FIG. 5 is a fragmentary vertical sectional view of a modified form
of the apparatus of FIG. 3.
Referring now to the drawings in detail, reference is first made to
FIGS. 1 and 2, wherein there is illustrated a laboratory growth
apparatus of this invention, generally designated by numeral
10.
The apparatus 10 includes a vessel 11, of glass construction, which
may be in the form of a twenty liter carboy, having a bottom wall
12, a main body or sidewall portion 13, an upper portion 14 of
reduced diameter relative to the body 13, and an upper rim 15
defining an opening therein, the vessel 11 being otherwise
imperforate. A glass cap 16, in the form of an inverted beaker,
having an end wall 17 and an annular sidewall 18, terminating at
its lower end in a rim 20 defining a generally circularly
configured hole or opening is provided, in inverted position
disposed about the neck portion 14 of the vessel 11.
An inoculation tube 21, also of glass construction is provided,
having a generally perpendicularly disposed major portion 22, and a
lower portion 23 angularly bent toward the lower corner of the
vessel 11 defined by the junction of the body wall 13 and bottom
wall 12, with the tube 21 having an open lower end 24. The upper
end of the tube 21 is reversely bent at 25, back over the vessel
rim 15, to be disposed downwardly between the vessel neck 14 and
the cap sidewall 18, in disposition close to the sidewall 18, with
the tube 21 being upwardly bent in the vicinity of the rim 20 of
the cap 16, to again have a vertically upwardly extending portion
27 which is fitted with a glass closure in the form of a serum cap
28. Reversely bent portion 26 of the glass tube 21 extends through
a slot 30 in the sidewall 18 of the cap 16, the slot 30 being cut
upwardly from the rim 20 of the cap 16, and being flared slightly
outwardly to be closely fitting about the tube portion 26, to
permit minimum air passage therethrough about the tube portion 26.
The tube 21 thus provides a means for inoculation of a culture 31
within the vessel 11. An additional glass tube 32 is provided,
virtually identical to the inoculation tube 21, but being used for
sampling, for example by extraction of a portion of the culture 31
by means of application of a vacuum to an upper portion 33 of the
glass tube 32, after removal of the serum cap 34 therefrom.
A third glass tube 35 is provided, generally configured similar to
the tubes 21 and 32, but having at its lower most end 36, inside
the vessel 11, a portion 37 in the form of a course porosity glass
frit 37, which provides a screening effect, as well as a diffusing
effect for air passing through the tube 35. An outer portion 38 of
the tube 35 is not bent upwardly as is, for example, the portion 27
of the tube 21, but is downwardly bent and is connected to an
enlarged glass universal joint 40, or ball joint, which contains a
cotton plug 41, several inches long, with the lower end of the
universal joint 40 also containing a cotton plug 42, to further
facilitate the filtering of particles from the air supply. A source
of a gas, such as sterile air is provided through the inlet 43 to
the ball joint 40, to enter into the culture 31 within the vessel
11 through the frit 37, to provide an atmosphere effective for
facilitating culture growth and the like. As this air bubbles up
through the culture 31, and into the upper portion of the interior
of the vessel 11, it continually escapes from the vessel 11, by
passing up over the rim 15 of the vessel 11, and downwardly between
the neck 14 of the vessel 11 and the sidewall 18 of the cap 16, to
pass outwardly of the apparatus 10 about the point of contact of
the rim 20 with the tapering or expanding portion of the body wall
13 of the vessel 11, as well as outwardly through the slots 30 in
the cap sidewall 18. Thus, this continuous discharge of air from
the apparatus 10 provides an effective seal preventing
contamination of the vessel 11 from an external source, at the
ambient pressure, in that the escaping pressure of the air from the
vessel 11 is at a greater pressure than the ambient pressure.
At the upper most end of the glass tube 32, just below the serum
cap 34, a small cotton plug (not shown) is disposed within the tube
32, held in place by small indentations in the glass, to provide a
means for removal or sampling through the tube 32. Similarly, the
tube 21 may be used for sampling, if such use is more desirable
than its use for inoculation, depending upon the particular culture
being studied, in which case, the tube 21 would also be provided
with a cotton plug 44 held in place by indentations in the glass
tube 21.
It is to be noted, that, while two such tubes 21 and 32 are
provided, either or both may be used for sampling and/or
inoculation, as desired, depending upon the particular
experiment.
The apparatus, for example the vessel 11 an the beaker 16 are
generally sterilized in an autoclave prior to use, to guard against
contamination, the tubes and other apparatus also being similarly
sterilized.
Stirring is provided by using a large diameter (for example 2 inch)
magnet 45 which has a coating 46 of glass, nylon or the like, to
render the same inert to the culture being studied, the magnet 45
being disposed approximately centrally of the vessel 11, at the
bottom thereof, and with the vessel 11 resting upon a magnet
actuating apparatus 47.
The magnetic stirring apparatus 47 includes a housing 48, a motor
50, which may be connected to the usual AC source, and a rotating
ferromagnet 51 driven by the motor 50, which, in turn, rotates the
magnetic element 45 within the vessel 11, for stirring the culture
31.
It is to be noted, in the apparatus mentioned above, the use of
ring seals is avoided, which works towards minimizing the cost of
the apparatus. Also, the use of seals on the vessel 11 is
avoided.
Referring now to FIGS. 3 and 4, an alternative apparatus 60 is
illustrated, for studies involving organisms which prefer a high
degree of aeration, such as lichen fungi. Because lichen fungi are
particularly notorious as being slow growers, the apparatus must
provide protection against contamination for long periods of time.
Also, provision must also be made for inoculation and removal of
solution and/or cell material, also with maximum protection against
contamination, as is required with the apparatus mentioned above.
The apparatus 60 utilizes a flask 61 which may be constructed in
the configuration of a one liter Erlenmeyer flask which has a pair
of tubes 62 and 63 in the form of sidearms (10 millimeter diameter
medium wall glass tubing) heat sealed thereto at 64 and 65,
respectfully, to communicate the interior bottom of the flask 61
with the outside. Suitable serum caps 66 and 66' are also provided.
In the tube 63, provision is made for removal of the solution with
filtration through a cotton plug 67, which is held in place by
glass indentations 68. The tube 63 would thus ordinarily be used
for removal of solution, and the tube 62 would be used for
inoculation.
A cap 70 is provided, having an annular sidewall portion 71 and an
end wall 72, for covering the open end 69 of the flask 61, seating
on the rim 74 thereof.
Sterile air is provided through the air inlet 75, which is filtered
through cotton plugs 76 and 77, in the ball joint 78, which then
passes into a glass tube 80 through an opening 81 in the end 72 of
the cap 70, with the tube 80 being connected in heat sealed
relation within the tube 72.
The tube 80 is angularly bent at 82 toward the periphery of the
bottom portion of the flask 61, and is also provided with a course
porosity frit 83.
A stirring element 84 which comprises a magnet having a Teflon,
glass or other suitable coating 85 is also provided in the bottom
of the flask 61, and is magnetically rotated by means of a rotating
ferromagnet 86 driven similarly the the magnet 51 of FIG. 1. The
device of FIG. 3 also operates in a manner similar to that of FIG.
1, in that it is the air passing through the culture 87, upwardly
outwardly around the rim 74 of the flask 61 and downwardly
outwardly of the cap 70 which provides a seal by means of the
pressure differential between the escaping air and the ambient air
pressure.
In the apparatus of FIGS. 3 and 4, the cotton plugs 76 and 77 also
provide a means for filtering particulate matter from the incoming
air. It will be noted, with respect to the air seal mentioned above
with respect to both apparatus, that the principal of the ordinary
petri dish is utilized, such that the outflow of air from the
apparatus tends to expel any foreign spores.
The apparatus has been used for the growth of lichen fungi, fungi,
and yeast, without visible evidence of contamination. These
apparatus have been used without contamination for periods as much
as several months. In some instances, as with fast growing fungi
such as those of the penicillin species, the inoculation may result
in such a heavy growth that the magnetic stirring becomes
ineffective in a matter of 2 or 3 days.
It will further be noted that, when the apparatus is used with
media having foaming tendencies, antifoams must be added to the
solutions 31 or 87, as necessary. However, in some instances, the
use of antifoams would not be desired, and accordingly, the
apparatus, particularly that of FIGS. 3 and 4 may be modified as
illustrated in FIG. 5, such that the tube 80 would extend
downwardly in a perpendicular manner, to a position directly above
the magnet 84, without being bent in the manner of FIG. 3, such
that the end of the tube 80 would be in fairly close proximity to
the magnet 84. By close control of this spacing and the size of the
magnet, as well as the rate of rotation of the magnet, a vortex
effect may be obtained which will create sufficient dispersion of
the incoming air into the solution 87.
It will be noted, that while two basic forms of the apparatus of
this invention have been disclosed in detail herein, various
modifications may be made in the structural details of the
apparatus of this invention, as well as in the use thereof, without
departing from the spirit and scope of the invention as defined in
the appended claims.
* * * * *