Apparatus and methods for automated diffusion filtration, culturing and photometric detection and enumeration of microbiological parameters in fluid samples

A method for analysis of microbiological parameters in a fluid sample, comprising the steps of:

• (a) providing a sterile sealed sample cuvette having a culture chamber and a detection chamber, the detection chamber being separated from the culture chamber by a permeable membrane, the permeable membrane having a pore size selected to block migration of a microbe under investigation from passing through the membrane into the detection chamber, each of the chambers having an inlet/outlet port for introduction, filtration and diffusion of fluid within the chambers;
• (b) pre-loading the culture camber with a reagent specific to the microbe under investigation, the reagent having a soluble optically active ingredient that provides a detectable parameter in the presence of the microbe;
• (c) introducing a fluid sample into the culture chamber through the inlet/outlet port using a suction filtration device and allowing the reagent to dissolve within the fluid sample to mix with reagent to form a dissolved reagent mixture;
• (d) enabling the fluid and the detectable parameter to migrate through the membrane into the detection chamber;
• (e) placing the sample cuvette into a holder within a housing and enclosing the housing;
• (f) incubating the sample by means of temperature control components mounted within the housing proximal to the holder for microbiological culturing;
• and
• (g) detecting the microbiological parameter by means of spectrophotometer components mounted within the housing proximal to the holder for photometric detection of the microbiological parameter.

A method for analysis of microbiological parameters in a fluid sample, comprising the steps of:

• (a) providing a sterile sealed sample cuvette having a culture chamber and a detection chamber, the detection chamber being separated from the culture chamber by a permeable membrane, the permeable membrane having a pore size selected to block migration of a microbe under investigation from passing through the membrane into the detection chamber, each of the chambers having an inlet/outlet port for introduction, filtration and diffusion of fluid within the chambers;
• (b) introducing a fluid sample into the culture chamber through the inlet/outlet port using a suction filtration device connected to the inlet/outlet port of the detection chamber;
• (c) filtering the sample by drawing the fluid through the membrane into the detection chamber and discarding the fluid through the outlet of the detection chamber until a desired volume of the sample is filtered;
• (d) introducing a reagent solution into the culture chamber, the reagent solution having a reagent specific to the microbe under investigation, the reagent having a soluble optically active ingredient that provides a detectable parameter in the presence of the microbe;
• (e) placing the sample cuvette into a holder within a housing and enclosing the housing;
• (f) incubating the sample by means of temperature control components mounted within the housing proximate the holder for microbiological culturing;
• and
• (g) detecting the microbiological parameter by means of spectrophotometer components mounted within the housing proximal to the holder for photometric detection of the microbiological parameter.