Techniques in Cytology – Cytospin: Cytospin Cell Resuspension Solution
In this blog series opening article, I discussed how cytospins could be used to separate cancer cells from non-cancer cells1. This cytology method …
Techniques in Cytology – Cytospin: Distinguishing Benign Cells from Malignant Cells
Cytology is a key component in diagnosis and screening of diseases such as cancer. It assesses single cells and clusters of cells from sources such …
Using Mass Spectrometry for Mass T cell Epitope Discovery
Time of Flight Mass Cytometry (CyTOF) is a relatively new multiparametric technology that is far outpacing standard fluorescence-based flow cytometry …
Whole Blood Phospho-flow: Direct Ex Vivo Measurement of Signaling in PBMC
I previously discussed phospho-flow cytometry, a method to study intracellular protein phosphorylation events in peripheral blood mononuclear cells …
Positive Selection vs Negative Selection for Cell Isolation
In a previous post, I covered the current options for isolating pure cell populations. One immediate question you will have to ask yourself is …
Optimizing Assays to Find Rare Antigen-Specific T cells in Cryopreserved PBMCs
Immunomonitoring of T cell based immune responses spans a wide range of therapeutic applications such as infectious and autoimmune diseases and is …
Highlight: How TNF knocks out Tregs!
A healthy and functional immune system requires a delicate balance of pro- and contra-inflammatory signals. Whereas, it is important to induce a …
Considerations for measuring cytokine levels in serum or plasma
Changes in circulating cytokine and chemokine levels have been associated with many human diseases, and thus understanding the relationships between …
Using Application Settings to Standardize Flow Cytometry Results Across Experiments and Instruments
While many fluorescence-based flow cytometry assays can be run without concern for hitting the exact same fluorescence intensity target values across …
The nuances of using CFSE to monitor lymphocyte proliferation
Measuring proliferation of lymphocytes such as T cells isolated from peripheral blood monuclear cells (PBMC) using carboxyfluorescein diacetate …
Identification of Type I Innate Lymphoid Cells that functionally resemble TH1 and NK cells
Innate lymphoid cells (ILCs) are subsets of lymphoid cells that do not rearrange their antigen receptors like T cells and B cells but have other …
Going Serum-Free in Cryopreserving PBMCs: Better Immunoassay Performance?
Probably the most common way to cryopreserve cells, including human peripheral blood mononuclear cells (PBMC) is using a mixture of 90% serum with …
Antigen Cross-Presentation by Human Dendritic Cell Subsets
Dendritic cells (DC) are major antigen-presenting cells consisting of numerous heterogeneous subtypes. In humans, several subtypes of DCs have …
Expansion of NK cells from Human PBMC
Natural killer (NK) cells represent up to 15% of human peripheral blood mononuclear cells (PBMC), and range from 5-20% of peripheral blood …
Are Terminally Differentiated Effector Memory Cells present in those “Naïve” CD4+ T cells you isolated from human PBMC?
Immunologists study many aspects regarding differentiation of T cells and function of T cell lineages. The results and interpretations from these …
Current Options for Isolating Pure Cell Populations
Antibody based isolation kits for isolating immune cell populations have become a standard protocol in the toolbox of every immunologist over the …
A New Subset of Negative Regulatory CD8 T Cells in Human PBMC
Negative regulatory CD4 T cells are well characterized and highly studied. However their CD8 counterparts are not well defined, particularly in …
Time of Flight Mass Cytometry (Cytof): Flying way beyond Fluorescent flow
Flow cytometry has been around since the 1950s when Wallace Coulter developed the first flow cytometry device and fluorescence-based flow cytometry …
Natural IgMs: A possible therapeutic role in autoimmunity?
Antibodies play a significant role in countering foreign threats and ensuring specificity of self/non-self recognition. However, in healthy …
Cool recent studies utilizing human PBMC
The availability of human peripheral blood mononuclear cells (PBMC) from healthy individuals and from patients with various diseases allows for many …
Artifacts and non-specific staining in flow cytometry, Part II
In Part I, I talked about un-specific binding and Fc-receptor binding. Besides these cases of non-specific binding, there are also other cases of …
Markers and functions of human CD4+ follicular helper T cells
CXCR5 is a chemokine receptor expressed by and used to identity human CD4+ follicular helper T cells (TFH). TFH cells, as their name implies, …
The Fascinating System of Eye-induced Immune Regulation
The immune privilege of the eye is a widely recognized but frequently oversimplified concept. The notion that the eye possessed unusual immunological …
Artifacts and non-specific staining in flow cytometry, Part I
If you add your antibody, lets say anti-CD3-epsilon antibody, to your cell solution you’d expect that only T cells will be labeled, right? …
Understanding MFI in the context of FACS data
Understanding MFI in the context of FACS data The speed, sensitivity and versatility of flow cytometry are things of beauty, but with great power …
Natural Killer Cell subtypes and markers in human PBMC
Natural Killer (NK) cells are a cytotoxic innate immune lymphocyte cell type. In humans, NK cells comprise up to 15% of peripheral blood …
Generation of CD4+ Th1 cells from human PBMC
CD4+ T helper type 1 (TH1) cells are the effector T cell population that governs cell mediated immune responses against intracellular pathogens …
Identification of CD8+ TC1, TC2, and TC17 populations in human PBMC
Human peripheral blood mononuclear cells (PBMC) are composed of heterogeneous populations of various immune cell types. CD4+ and CD8+ T cells are …