ArticleThe journal of histochemistry and cytochemistry : official journal of the Histochemistry Society2020
A Sample Preparation Protocol for High Throughput Immunofluorescence of Suspension Cells on an Adherent Surface.
Article in The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society, 2020. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 10 papers.
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Who cites it
10 citing papers in PubMed, 17 citations in OpenAlex.
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- Effects of ex vivo blood anticoagulation and preanalytical processing time on the proteome content of platelets.Journal of thrombosis and haemostasis : JTH · 2022Article
- HiIDDD: a high-throughput imaging pipeline for the quantitative detection of DNA damage in primary human immune cells.Scientific reports · 2022Article
- Double Labeling Fluorescent Immunocytochemistry.Methods in molecular biology (Clifton, N.J.) · 2022Article
- Investigating the Impact of the Bit Depth of Fluorescence-Stained Images on the Performance of Deep Learning-Based Nuclei Instance Segmentation.Diagnostics (Basel, Switzerland) · 2021Article
- Platelet Proteomes, Pathways, and Phenotypes as Informants of Vascular Wellness and Disease.Arteriosclerosis, thrombosis, and vascular biology · 2021Review
- Transient Destabilization of Biological Membranes Contributes to the Superior Performance of Star-Shaped PDMAEMA in Delivering pDNA.ACS omega · 2020Article
- Serum-Independent Nonviral Gene Delivery to Innate and Adaptive Immune Cells Using Immunoplexes.ACS applied bio materials · 2020Article
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Authors and funding
7 authors at 2 institutions in 2 countries.
Funding
Abstract
Imaging is a powerful approach for studying protein expression and has the advantage over other methodologies in providing spatial information in situ at single cell level. Using immunofluorescence and confocal microscopy, detailed information of subcellular distribution of proteins can be obtained. While adherent cells of different tissue origin are relatively easy to prepare for imaging applications, non-adherent cells from hematopoietic origin, present a challenge due to their poor attachment to surfaces and subsequent loss of a substantial fraction of the cells. Still, these cell types represent an important part of the human proteome and express genes that are not expressed in adherent cell types. In the era of cell mapping efforts, overcoming the challenge with suspension cells for imaging applications would enable systematic profiling of hematopoietic cells. In this work, we successfully established an immunofluorescence protocol for preparation of suspension cell lines, peripheral blood mononucleated cells (PBMC) and human platelets on an adherent surface. The protocol is based on a multi-well plate format with automated sample preparation, allowing for robust high throughput imaging applications. In combination with confocal microscopy, the protocol enables systematic exploration of protein localization to all major subcellular structures.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.