Determining Cell Number During Cell Culture using the Scepter Cell Counter

Abstract

Counting cells is often a necessary but tedious step for in vitro cell culture. Consistent cell concentrations ensure experimental reproducibility and accuracy. Cell counts are important for monitoring cell health and proliferation rate, assessing immortalization or transformation, seeding cells for subsequent experiments, transfection or infection, and preparing for cell-based assays. It is important that cell counts be accurate, consistent, and fast, particularly for quantitative measurements of cellular responses.Despite this need for speed and accuracy in cell counting, 71% of 400 researchers surveyed 1 who count cells using a hemocytometer. While hemocytometry is inexpensive, it is laborious and subject to user bias and misuse, which results in inaccurate counts. Hemocytometers are made of special optical glass on which cell suspensions are loaded in specified volumes and counted under a microscope. Sources of errors in hemocytometry include: uneven cell distribution in the sample, too many or too few cells in the sample, subjective decisions as to whether a given cell falls within the defined counting area, contamination of the hemocytometer, user-to-user variation, and variation of hemocytometer filling rate 2 .To alleviate the tedium associated with manual counting, 29% of researchers count cells using automated cell counting devices; these include vision-based counters, systems that detect cells using the Coulter principle, or flow cytometry 1 . For most researchers, the main barrier to using an automated system is the price associated with these large benchtop instruments 1 .The Scepter cell counter is an automated handheld device that offers the automation and accuracy of Coulter counting at a relatively low cost. The system employs the Coulter principle of impedance-based particle detection 3 in a miniaturized format using a combination of analog and digital hardware for sensing, signal processing, data storage, and graphical display. The disposable tip is engineered with a microfabricated, cellsensing zone that enables discrimination by cell size and cell volume at sub-micron and sub-picoliter resolution. Enhanced with precision liquid-handling channels and electronics, the Scepter cell counter reports cell population statistics graphically displayed as a histogram. Video LinkThe video component of this article can be found at

Facile generation of giant unilamellar vesicles using polyacrylamide gels

“…A Scepter ™ 2.0 Handheld Cell Counter (EMD Millipore) was used to quantify the mean diameter and concentration of the GUVs. This device, traditionally used in mammalian cell culture, is based on the Coulter Principle; as each cell passes through an aperture, a voltage spike is recorded and its magnitude is correlated to particle size 46,47 . 60 µm tips were used, which are capable of detecting particles between 6 and 36 µm.…”

A high-throughput AO/PI-based cell concentration and viability detection method using the Celigo image cytometry

“…Cell concentration and viability are two simple yet important parameters to measure when working with cell-based research such as immuno-oncology, toxicology, and bioprocessing (Ongena et al 2010;Chan et al 2011a, b;2012b). Assays from standard cell culture, plating cells for ELISA, to preparing cells for flow cytometric analysis, all require accurate measurements of cell concentration and viability in order to generate meaningful and comparable results (Chan et al 2013).…”

Rapid and Sensitive Quantitation of Heme in Hemoglobinized Cells

“…MEL and K562 cells were washed and diluted in 1 PBS before cell counts were done with a Scepter handheld cell counter (Millipore, Billerica, MA) as previously described (22). MEL and K562 cell proliferation values were obtained by dividing the ratio of live cells at the end of each time course to the number of cells seeded at the beginning by the same ratio for untreated cultures.…”