Methods of collecting cells from multi-well plates for use in flow cytometry

Abstract

A method of collecting cells from individual wells of a multi-well plate for flow cytometry, the method comprising: determining particle or cell counts from each well of the multi-well plate by flow cytometry; ordering the wells from lowest particle count to highest particle count; generating a sequential order for collection that follows the ordering of wells to establish a collection pattern; and collecting particles or cells according to the collection pattern in a second multi-well plate by flow cytometry.

What is claimed is: 
     
         1 . A method of collecting cells from individual wells of a multi-well plate for flow cytometry, the method comprising:
 determining particle counts from each well of the multi-well plate by flow cytometry;   ordering the wells from lowest particle count to highest particle count;   generating a sequential order for collection from the ordering of wells to establish a collection pattern; and   collecting particles according to the collection pattern in a second multi-well plate by flow cytometry.   
     
     
         2 . The method of  claim 1 , wherein the particle counts are a number of counted cells and the collected particles are collected cells. 
     
     
         3 . The method according to  claim 1 , wherein the multi-well plate is a 96 well plate, characterized as having rows A-H and columns 1-12, further wherein the middle region is characterized as a well selected from the group consisting of wells D6, D7, E6 and E7. 
     
     
         4 . The method  claim 1 , wherein the step of ordering the wells comprises grouping the wells by count ranges. 
     
     
         5 . The method  claim 4 , wherein the sequential order is such that groups of a lower count range are ordered before groups of a higher count range. 
     
     
         6 . The method of  claim 5 , wherein the sequential order of wells within each group follow a same pattern. 
     
     
         7 . The method according to  claim 1 , wherein the step of generating the sequential order comprises establishing a successive square pattern or a spiral square pattern starting from the well having the lowest particle count or from a well within a group of wells having a lowest count range. 
     
     
         8 . The method according to  claim 1 , wherein the collection pattern exactly follows the ordering of wells from lowest particle count to highest particle count. 
     
     
         9 . The method according to  claim 1 , wherein the collection pattern deviates from the ordering of wells less than 15%. 
     
     
         10 . The method according to  claim 1 , wherein the method further includes rotating or agitating the multi-well plate between steps of collecting particles from different wells. 
     
     
         11 . The method according to  claim 1 , wherein the method further comprises a second collection pattern performed after the collection pattern. 
     
     
         12 . The method according to  claim 1 , wherein the second collection pattern is a meandering pattern between two columns of wells. 
     
     
         13 . The method according to  claim 1 , further comprising labeling the cells with labelled binding reagents against one or more cell biomarkers. 
     
     
         14 . The method according to  claim 13 , wherein the labelled binding reagents are fluorescently labelled antibodies or antibody fragments.