Cell Suspension Pooling
Cell Suspension Pooling - Related Videos
09:12
MISSION LentiPlex Pooled shRNA Library Screening in Mammalian Cells
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Cited by 10 •
2011
Here we use a human LentiPlex pooled library and traditional sequencing methods to identify gene targets promoting cell survival. We demonstrate how to set up and deconvolute a LentiPlex screen and validate the results.
08:32
Preparation of Pooled Human Platelet Lysate (pHPL) as an Efficient Supplement for Animal Serum-Free Human Stem Cell Cultures
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Cited by 133 •
2009
Human platelet lysate is a rich source of growth factors and a potent supplement in cell culture. This protocol presents the process of preparing a large pool of human platelet lysate by starting from platelet rich plasma, performing several freeze-thaw cycles and depleting the platelet fragments.
00:09
Pooled CRISPR-Based Genetic Screens in Mammalian Cells
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Cited by 13 •
2019
CRISPR-Cas9 technology provides an efficient method to precisely edit the mammalian genome in any cell type and represents a novel means to perform genome-wide genetic screens. A detailed protocol discussing the steps required for the successful performance of pooled genome-wide CRISPR-Cas9 screens is provided here.
11:17
Generation of Single-Cell Suspensions from Mouse Neural Tissue
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Cited by 16 •
2009
Dissociating cells from specific tissue types requires specific parameters for tissue aggitation to obtain a high volume of viable, culturable cells. The Miltenyi gentleMACS Dissociator optimizes this task with a simple, practical protocol. In this publication the use of this apparatus on nerual tissue is explained.
13:55
Measurement of Vacuolar and Cytosolic pH In Vivo in Yeast Cell Suspensions
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Cited by 36 •
2013
Vacuolar and cytosolic pH can be measured in live yeast (S. cerevisiae) cells using ratiometric fluorescent dyes localized to specific cellular compartments. We describe procedures for measuring vacuolar pH with BCECF-AM, which localizes to the vacuole in yeast, and cytosolic pH with a cytosolic ratiometric pH-sensitive GFP (yeast pHluorin).
FAQs
Gentle resuspension helps distribute cells throughout the combined preparation without unnecessarily compromising viability. A uniform mixture is important because a sampled aliquot should more consistently represent the pooled material. In biological workflows, this supports dependable cell counting, flow-cytometry measurements, culture setup, and molecular profiling by reducing variation caused by uneven cell distribution.
Cell concentration and viability determine how consistently the pooled material can serve as downstream input. Controlling concentration helps researchers compare or standardize the amount of cellular material used, while monitoring viability indicates whether the preparation remains suitable for subsequent work. Together, these measures improve experimental consistency and support more reliable biological comparisons.
Sample identity must remain controlled throughout pooling so researchers can interpret the resulting preparation correctly. This is especially important when combining biological replicates or preparing material for comparative analyses. Clear identity control, together with defined contributions from each suspension, helps preserve the intended experimental design and reduces uncertainty when results are evaluated.
Researchers collect defined volumes or cell numbers from the suspensions before combining them. This planned input allows the pooled preparation to be assembled consistently and can help standardize the material supplied to a downstream assay. Selecting contributions in advance also supports controlled consolidation of biological replicates rather than relying on unmeasured additions.
A basic workflow includes collecting the planned volume or cell number from each suspension, combining the material under sterile conditions, and gently resuspending it to promote even distribution. Researchers then control relevant characteristics such as concentration, viability, and sample identity before using the preparation. These steps support consistent handling across laboratory workflows.
Pooling is useful when researchers need to consolidate biological replicates, standardize downstream input, or obtain enough material for a planned assay. The resulting preparation can support cell counting, flow cytometry, culture, and molecular profiling. Its value depends on maintaining controlled inputs and suitable mixing so the material remains consistent for the selected application.