US 11,939,621 B2
Device for determining live/dead bacterial state and method for determining live/dead bacterial state using the device
Toshikazu Moriwaki, Fukuoka (JP); and Hideki Moriwaki, Fukuoka (JP)
Assigned to NIHON RIKAGAKU KAIHATSU LLC., Fukuoka (JP)
Filed by NIHON RIKAGAKU KAIHATSU LLC., Fukuoka (JP)
Filed on Sep. 7, 2021, as Appl. No. 17/468,617.
Application 17/468,617 is a continuation of application No. 15/778,877, granted, now 11,118,209, previously published as PCT/JP2016/088798, filed on Dec. 26, 2016.
Claims priority of application No. 2015-257250 (JP), filed on Dec. 28, 2015.
Prior Publication US 2021/0403974 A1, Dec. 30, 2021
This patent is subject to a terminal disclaimer.
Int. Cl. C12M 1/00 (2006.01); C12M 1/12 (2006.01); C12M 1/34 (2006.01); C12Q 1/00 (2006.01); C12Q 1/02 (2006.01); C12Q 1/06 (2006.01)
CPC C12Q 1/06 (2013.01) [C12M 1/12 (2013.01); C12M 1/34 (2013.01); C12Q 1/02 (2013.01)] 2 Claims
OG exemplary drawing
 
1. A method for determining a live/dead bacterial state of bacteria by utilizing a determination device for determining the live/dead bacterial state comprising a step for filtering and collecting the bacteria on a measurement filter,
wherein the determination device for determining the live/dead bacterial state comprises:
a case body inside of which is a darkroom space;
an opening/closing lid body which opens and closes a bacteria base insertion port formed in front of the case body;
a measurement mechanism housed in the case body and configured to determine a live bacterial state and a dead bacterial state of the bacteria collected from a specimen and to measure a number of bacteria;
wherein the measurement mechanism comprises:
a bacteria base holding mechanism that holds a bacteria base on which the bacteria collected on the measurement filter from the specimen is placed;
an excitation light irradiation mechanism arranged to irradiate light toward the bacteria on the bacteria base;
an imaging camera arranged to image the bacteria on the bacteria base;
an XY-axes adjustment mechanism for moving and adjusting an XY stage having two separately moving layers and supporting the bacteria base holding mechanism in an XY-axes direction,
wherein the bacteria base holding mechanism comprises:
left and right rails for clamping both side edges of the bacteria base from left and right outer side directions;
a rail displacement mechanism configured to displace one of the left and right rails toward outside and release the clamping of the bacteria base by the left and right rails; and
an operation lever arranged outside of the one of the left and right rails to operate the rail displacement mechanism from the bacteria base insertion port and the operation lever rotationally operate around a support positioned midway of the operation lever,
wherein the rail displacement mechanism comprises:
a slide shaft horizontally projected on the one of the left and right rails;
an operation lever contact projection formed on a head portion of the slide shaft;
an operation support portion supporting a midway portion of the operation lever, a top portion of the operation lever contacts the operation lever contact projection on the XY stage; and
a compression spring provided between an outer side surface of the one of the left and right rails and the top portion of the operation lever, and
wherein the excitation light irradiation mechanism has an LED arranged to irradiate the bacteria with excitation light to excite a fluorescent substance which was used to dye the bacteria so as to respectively detect a live bacterial state and a dead bacterial state,
wherein the filtering and collecting the bacteria on the measurement filter comprises:
dropping the specimen in a test tube;
dropping a reagent containing the fluorescent substance in the test tube and sealing and standing the test tube still after stirring;
housing and setting the measurement filter in a filter housing;
connecting the filter housing in which the measurement filter is housed to a top portion of a syringe and forming a communication state between an inner side of the filter housing and an inner side of the syringe;
injecting the specimen in the test tube into the inner side of the syringe;
pressing the inner side of the syringe and pushing the bacteria together with the specimen toward the measurement filter in the filter housing from the top portion of the syringe, thereby filtering and collecting the bacteria stained on a surface of the measurement filter;
taking out the measurement filter from the filter housing and putting the measurement filter on the bacteria base, thereby arranging the bacteria filtered and collected on the surface of the measurement filter on the bacteria base through the measurement filter; and
inserting the bacteria base in the bacteria base holding mechanism provided in the determination device for determining the live/dead bacterial state ands determining the live/dead bacterial state by utilizing the excitation light irradiation mechanism and the imaging camera provided in the determination device and grasping a bacteria state as images.