U.S. patent number 3,630,849 [Application Number 04/820,040] was granted by the patent office on 1971-12-28 for surface micro-organism contamination assays.
Invention is credited to Stephen L. Bazil, David B. Land.
United States Patent |
3,630,849 |
Land , et al. |
December 28, 1971 |
SURFACE MICRO-ORGANISM CONTAMINATION ASSAYS
Abstract
A receptacle and lid combination for molding solidified nutrient
containing agar-agar with a flat face coplanar with the rim of the
receptacle and keyed in the receptacle to resist withdrawal, for
micro-organism contamination assays. The lid is formed with a flat
base inside and the height of the circular wall on the receptacle
is greater than the height of the circular wall on the lid to
permit the receptacle to seat against the base of the lid and where
the receptacle is formed with a vent and the lid is formed with
projections from the wall near the base for a force fit with the
receptacle.
Inventors: |
Land; David B. (Flushing,
NY), Bazil; Stephen L. (Valley Stream, NY) |
Family
ID: |
25229724 |
Appl.
No.: |
04/820,040 |
Filed: |
April 24, 1969 |
Current U.S.
Class: |
435/305.4 |
Current CPC
Class: |
C12M
41/36 (20130101); C12M 23/10 (20130101); C12M
23/38 (20130101) |
Current International
Class: |
C12M
1/22 (20060101); C12k 001/00 () |
Field of
Search: |
;195/103.5,139LE,222
(189)/ ;195/222 (540)/ ;195/222 (460)/ ;195/220 (44/ A)/ |
References Cited
[Referenced By]
U.S. Patent Documents
Foreign Patent Documents
Primary Examiner: Monacell; A. Louis
Assistant Examiner: Hensley; Max D.
Claims
We claim:
1. A receptacle and lid combination for nutrient-containing
agar-agar for use in monitoring for micro-organisms comprising
a lid having a base with an essentially flat inside surface and a
circular wall with at least three short equal height projections
extending inwardly from the circular wall near said base, and
a receptacle having a base and a circular wall extending from the
base of the receptacle and slightly longer than the circular wall
of the lid and of outside diameter to fit into the circular wall of
the lid to the projections and to force fit within the projections
to seat against the base of the lid said base of said receptacle
having a vent hole.
2. A receptacle and lid combination as defined in claim 1
wherein
the circular wall of the receptacle is frustoconical and has its
large diameter end at the base,
the circular wall of the lid is frustoconical and has its small
diameter end at the base, and
the short equal height projections project inwardly from the wall
at the base.
3. A receptacle and lid combination as defined in claim 1 wherein
the circular wall of the receptacle is formed with at least one
arcuate inwardly directed projection for keying agar-agar that may
be solidified in the receptacle.
4. A receptacle and lid combination as defined in claim 2 wherein
the arcuately inwardly directed projection is a helical turn.
5. A receptacle and lid combination as defined in claim 2 wherein
the circular wall of the receptacle is formed with four identical
arcuate inwardly directed projections.
6. A receptacle and lid combination as defined in claim 1 wherein
the center of the base of the receptacle is formed with a
knockout.
7. A receptacle and lid combination as defined in claim 1 wherein
the center of the base of the receptacle is formed with a coaxial
cylinder for nesting a fibrous plug to permit passage of air but
serving as a barrier to micro-organisms.
8. A receptacle and lid combination for nutrient containing
agar-agar for use in monitoring for micro-organisms comprising
a lid having a base with an essentially flat inside surface and a
frustoconical circular wall, and
a receptacle having a base and a frustoconical circular wall having
its large diameter end at the base of the receptacle extending from
the base of the receptacle and slightly longer than the circular
wall of the lid and of outside diameter to fit into the circular
wall of the lid to seat against the base of the lid, the base of
the receptacle increasing in thickness from the center outwardly
toward the circular wall said base of said receptacle having a vent
hole.
Description
The invention described herein may be manufactured and used by or
for the Government of the United States of America for governmental
purposes without the payment of any royalties thereon or
therefor.
BACKGROUND OF THE INVENTION
A technique for monitoring the quantity of micro-organisms that
settle from the atmosphere in any particular outdoor or indoor
location is to place at that location an upwardly directed open
receptacle with shallow depth sterile solidified
nutrient-containing clear agar-agar. The receptacle is transparent
and is formed with a grid on the bottom delineating square
centimeters. The depth of agar-agar is on the order of one-fourth
inch. The period of exposure to the atmosphere is on the order of
12-24 hours. Micro-organisms that have deposited during that time
cannot be seen even if the transparent receptacle and agar-agar is
brightly illuminated. The receptacle is covered and is placed in an
incubator set at temperature within the range 25.degree. to
37.degree. C. for about a day. The agar-agar remains solid during
incubation; it liquifies at 60.degree. C. and solidifies at
45.degree. C. Individual micro-organisms captured on the surface of
the agar-agar that thrive on the nutrient multiply into clusters or
colonies during incubation. After the incubation period, the
receptacle is illuminated from the rear. The colonies appear as
dark spots to the naked eye. The assay is carried out by counting
the number of dark spots in one or more of the squares.
A preferred modification of the above described technique that has
been in common use eliminates the step of leaving an upwardly
directed open receptacle in situ for hours and instead involves
butting a surface of a thin agar-agar disk against a surface in a
location to be monitored and withdrawing the agar-agar.
Micro-organisms and minute particles on the monitored surface
adhere to the agar-agar. The steps of culturing and assaying are
otherwise the same as described above. The agar-agar disk for the
modified technique is molded in a shallow transparent receptacle
which is on the order of 2 inches in diameter and between
one-eighth and one-fourth inch deep with centimeter square
delineations in a grid pattern. A commercially marketed receptacle
for the technique described is RODAC PLATE, a registered trademark
of Falcon Plastics. Though termed a plate, this type product
includes a shallow receptacle and a lid for that receptacle. This
plate is designed to nest in another plate whereby several of them
can be stacked in an incubator for space economy. In use, the
shallow receptacle is filled with nutrient-containing agar-agar.
The agar-agar is permitted to solidify with the lid in place. The
surface of the solidified agar-agar is not coplanar with the rim;
it has a concave meniscus. The lidded receptacle containing the
solidified agar-agar is brought to a flat surface to be monitored,
the lid is removed, the receptacle is overturned and because of the
meniscus needs to be pressed to flatten the concave surface of the
agar-agar to force the face against the surface to be monitored. A
percentage of the receptacles are ruined because the polystyrene or
other resin of which the receptacle is formed cracks. In some other
cases the agar-agar does not release from the monitor surface and
instead is partly or completely pulled free of the receptacle when
the receptacle is withdrawn. Because the pressure applied cannot be
uniform and of the same magnitude for each disk, a variable is
introduced limiting precision of the technique. Also this type of
product can be used for growing aerobic micro-organisms only.
An object of this invention is to provide a receptacle and lid
combination of the type described that overcomes the objections of
the prior art, that is inexpensive, timesaving, takes up little
room for growing aerobic and anaerobic bacteria and can be used for
accurately determining surface contamination in hospitals, clinical
laboratories, bacteriological research institutions,
food-processing areas and food consumption areas, or any geographic
area of BW defense program.
Other objects, advantages and novel features of the invention will
become apparent from the following detailed description of the
invention when considered in conjunction with the accompanying
drawings wherein:
FIG. 1 is a top plan view of a prior art receptacle,
FIG. 2 is a cross section of the receptacle of FIG. 1 with the lid
in place,
FIG. 3 is a plan view of a receptacle in accordance with the
principles of this invention,
FIG. 4 is sectional view of the receptacle shown in FIG. 3 with the
lid in place,
FIG. 5 is a perspective view of another embodiment of
receptacle,
FIG. 6 is a cross-sectional view of another embodiment of
receptacle and lid combination according to the principles of this
invention, and
FIG. 7 shows a structural arrangement of vent and handle for any of
the receptacles.
SUMMARY OF THE INVENTION
An agar-agar receptacle is formed to key the solidified agar-agar
against withdrawal when the receptacle is inverted. The agar-agar
is solidified in the receptacle while inverted and seated on a flat
surface or otherwise selectively shaped surface to form a flat or
selected shape agar-agar face .
DESCRIPTION
There is shown in FIGS. 1 and 2 a prior art type of receptacle 10
and lid 12 combination for monitoring surface areas for
micro-organisms. The receptacle has a circular base with a short
skirt 14 projecting from one face at the perimeter and a short
circular ridge 16 of lesser diameter than the base projecting from
the opposite face of the base. The lid 12 is circular, of larger
diameter than the ridge 16 and deeper than the ridge to contain an
air space and seats on the base around the ridge and of lesser
diameter than the skirt for nesting into the skirt of another
receptacle for stacking purposes. Several short projections are
formed on the outer face of the lid whereby when the units are
stacked in an incubator there is air circulation for uniform
temperature.
In FIGS. 3 and 4 there is shown an embodiment of a receptacle 20
and lid 22 according to this invention. The receptacle includes a
flat base 24 and a circular wall 26 projecting from one face of the
base 24 and of smaller diameter than the base. Thin arcuate
inwardly directed fins 28 project from the wall parallel to the
base intermediate the base and the free end of the wall 26. The
base 24 has a knockout 30. The lid 22 has a base 32 with an inner
flat face and formed with a grid pattern of centimeter squares and
a circular wall 34 slightly greater in diameter than the diameter
of wall 26. The height of the wall 34 measured from the base 32 is
slightly less than the height of wall 26 measured from the base 24.
Three small integral nipplelike projections 36 are directed
inwardly from the wall 34 near the base 32 and are of a size such
that the lid and receptacle must be forced together for the
receptacle to seat against the base 32 of the lid. If not forced
together, the receptacle rests on the projections 36 when the lid
22 is underneath. The combination of receptacle and lid are
packaged sterile.
In use, the combination shown in FIG. 4 is removed from the
packaging and knockout 30 is displaced. The lid is substantially
filled with nutrient-containing agar-agar and the receptacle is
forced into the lid to seat against the bottom. An adhesive tape is
used to cover the opening in the receptacle for shielding against
contamination. The agar-agar is permitted to solidify. Because the
free face of the agar-agar is solidified against a flat surface,
the face is flat and coplanar with the rim of the receptacle. The
receptacle and lid combination is taken to the surface to be
monitored, separated, and the receptacle is butted against the
surface. Since the face of the agar-agar is flat, the entire face
contacts the surface being monitored without pressure. If the
surface to be monitored has a special shape, the base of the lid is
made with a corresponding geometry. Then the receptacle is nested
again in the lid and placed in the incubator. To culture anaerobic
micro-organisms adhered to the agar-agar surface and not the
aerobic micro-organisms, the receptacle and lid are pressed
together, forcing out essentially all the air from between the face
of the agar-agar and the inner face of the lid. Alternatively, the
receptacle is nested in the lid on the nipplelike projections
leaving an air space and permitting access of the atmosphere to the
face of the agar-agar. After the incubation time, if the receptacle
and lid were not previously pressed together, they are then pressed
together. The combination is looked through toward a bright source.
Spots per square in one or more of the squares are counted.
FIG. 5 shows another form of receptacle 38 which differs from the
receptacle 20 in FIG. 3 in that it includes a helical one-turn
keying projection 40 instead of the keying projections 28 but is
otherwise used in the same manner as is described. When molded, the
receptacle 38 is unscrewed for removal; the mold for making the
receptacle shown in FIG. 3 is more expensive.
The embodiment shown in FIG. 6 includes receptacle and lid 42 and
44 wherein the circular wall of each is frustoconical to serve the
same purpose as the keying projections 28 in FIG. 3 and the keying
projection 40 in FIG. 5. Instead of projection 36 in FIG. 4,
projections 46 extend from the wall at the base of the lid. The
receptacle 42 may be molded in one piece or may be made by bonding
separate base and wall portions and then cementing or fusing
them.
Each of the described receptacles may be formed with a knockout as
described above, or with a hole that is sealed on the outer side
with adhesive tape or with coaxial central cylindrical projection
50 for a fibrous plug 52 as shown in FIG. 7 for permitting passage
of air and serving as a barrier against micro-organisms.
Additionally, a handle 54 may be provided on the outer side of the
receptacle.
Another feature included in this invention not shown on the
drawings is to form the base of the receptacle so that the outer
surface is flat and that the inner surface is slightly domed or
coned. In other words the thickness of the base increases from its
center outwardly to the circular wall of the receptacle. The
purpose is to prevent the occlusion of bubbles of air around the
inner surface of the base. By virtue of the domed or coned shape
surface the air bubbles are driven to the center opening and do not
occlude along the inner surface of the base.
* * * * *