ASTM F2149-01(2007)
Standard Test Method for Automated Analyses of Cells-the Electrical Sensing Zone Method of Enumerating and Sizing Single Cell Suspensions

Standard No.

ASTM F2149-01(2007)

Release Date

2001

Published By

American Society for Testing and Materials (ASTM)

Status

Be replaced

Replace By

ASTM F2149-16

Latest

ASTM F2149-16

Scope

This assay is used in university tissue culture laboratories, government research, and hospital, biomedical, and pharmaceutical laboratories to automate cell counting and sizing. This instrumentation provides very rapid, accurate, and precise results for any tissue culture facility. In addition, as noted, since the cell sizes to be analyzed by the instrument are set by the user, the analyses may be done on virtually any species of cells and cell type; it is not restricted to human cells or blood cells.

The electrical sensing zone methodology was introduced in the mid 1950s (9). Since this time, there have been substantial improvements which have enhanced the operatorrsquo;ease of use. Among these are the elimination of the mercury manometer, reduced size, greater automation, and availability of comprehensive statistical computer programs.

This instrumentation offers a rapid result as contrasted to the manual counting of cells using the standard counting chamber, hemocytometer. The counting chamber is known to have an error of 10 to 30 %, as well as being very time consuming (10). In addition, when counting and sizing porcine hepatocytes, Stegemann et al concluded that the automated, electrical sensing zone method provided significantly greater accuracy, precision, and speed, for both counts and size, compared to the conventional microscopic or the cell mass-based method (7).

1.1 This test method, provided the limitations are understood, covers a procedure for both the enumeration and measurement of size distribution of most all cell types. The instrumentation allows for user-selectable cell size settings, hence, this test method is not restricted to specific cell types. The method is appropriate for suspension as well as adherent cell cultures (). This is a quantitative laboratory method not intended for on-line or field use. Results may be reported as number of cells per millilitre or total number of cells per volume of cell suspension analyzed. Both count and size distribution may be expressed in cell micron diameter or volume, femtolitres.

1.2 Cells commonly used in tissue-engineered medical products () routinely are analyzed. Examples are chondrocytes (), fibroblasts (), and keratinocytes (). Szabo et al used the method for both pancreatic islet number and volume measurements (). In addition, instrumentation using the electrical sensing zone technology was used for both count and size distribution analyses of porcine hepatocytes placed into hollow fiber cartridge extracorporeal liver assist systems. In this study (), and others (, ), the automated electrical sensing zone method was clearly validated for superior accuracy and precision when compared to the conventional manual method, visual cell counting under a microscope using a hemocytometer. This validation has been demonstrated over a wide variety of cell types. In addition, the automated procedure is rapid, rugged, and cost effective; it also minimizes operator-to-operator variability inherent in manual techniques.

1.3 This instrumentation is manufactured by a variety of companies; however, the principle used in all is electrical impedance. This test method, for cell counting and sizing, is based on the detection and measurement of changes in electrical resistance produced by a cell, suspended in a conductive liquid, traversing through a small aperture (see ()). When cells are suspended in a conductive liquid, phosphate-buffered saline for instance, they function as discrete insulators. When the cell suspension is drawn through a small cylindrical aperture, the passage of each cell changes the impedance of the electrical path between two submerged electrodes located on each side of the aperture. ......

ASTM F2149-01(2007) history

• 2016 ASTM F2149-16 Standard Test Method for Automated Analyses of Cells—the Electrical Sensing Zone Method of Enumerating and Sizing Single Cell Suspensions
• 2001 ASTM F2149-01(2007) Standard Test Method for Automated Analyses of Cells-the Electrical Sensing Zone Method of Enumerating and Sizing Single Cell Suspensions
• 2001 ASTM F2149-01 Standard Test Method for Automated Analyses of Cells8212;the Electrical Sensing Zone Method of Enumerating and Sizing Single Cell Suspensions