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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 …

August 20, 2013 | PBMC Basics | 0 Comments
Techniques in Cytology – Cytospin: Distinguishing Benign Cells from Malignant Cells

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 …

July 31, 2013 | PBMC Basics | 0 Comments

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 …

July 15, 2013 | Immunology, PBMC Basics | 0 Comments
Whole Blood Phospho-flow: Direct Ex Vivo Measurement of Signaling in PBMC

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 …

July 10, 2013 | Immunology, PBMC Basics, Protocols | 0 Comments
Positive Selection vs Negative Selection for Cell Isolation

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 …

June 18, 2013 | PBMC Basics | 0 Comments
Optimizing Assays to Find Rare Antigen-Specific T cells in Cryopreserved PBMCs

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 …

May 31, 2013 | PBMC Basics | 0 Comments
Highlight: How TNF knocks out Tregs!

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 …

May 30, 2013 | PBMC Basics | 0 Comments
Considerations for measuring cytokine levels in serum or plasma

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 …

May 28, 2013 | PBMC Basics | 0 Comments
Using Application Settings to Standardize Flow Cytometry Results Across Experiments and Instruments

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 …

May 17, 2013 | PBMC Basics | 0 Comments
The nuances of using CFSE to monitor lymphocyte proliferation

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 …

May 9, 2013 | PBMC Basics | 0 Comments
Identification of Type I Innate Lymphoid Cells that functionally resemble TH1 and NK cells

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 …

April 29, 2013 | PBMC Basics | 2 Comments
Going Serum-Free in Cryopreserving PBMCs: Better Immunoassay Performance?

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 …

April 22, 2013 | PBMC Basics | 0 Comments
Antigen Cross-Presentation by Human Dendritic Cell Subsets

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 …

April 12, 2013 | PBMC Basics | 0 Comments
Expansion of NK cells from Human PBMC

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 …

April 8, 2013 | PBMC Basics | 0 Comments
Are Terminally Differentiated Effector Memory Cells present in those “Naïve” CD4+ T cells you isolated from human PBMC?

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 …

April 5, 2013 | PBMC Basics | 1 Comment
Current Options for Isolating Pure Cell Populations

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 …

April 4, 2013 | PBMC Basics | 0 Comments
A New Subset of Negative Regulatory CD8 T Cells in Human PBMC

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 …

April 3, 2013 | PBMC Basics | 0 Comments
Time of Flight Mass Cytometry (Cytof): Flying way beyond Fluorescent flow

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 …

April 1, 2013 | PBMC Basics | 0 Comments
Natural IgMs: A possible therapeutic role in autoimmunity?

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 …

March 29, 2013 | PBMC Basics | 0 Comments
Cool recent studies utilizing human PBMC

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 …

March 29, 2013 | PBMC Basics | 0 Comments
Artifacts and non-specific staining in flow cytometry, Part II

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 …

March 26, 2013 | PBMC Basics | 0 Comments
Markers and functions of human CD4+ follicular helper T cells

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, …

March 21, 2013 | PBMC Basics | 0 Comments
The Fascinating System of Eye-induced Immune Regulation

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 …

March 21, 2013 | PBMC Basics | 0 Comments
Artifacts and non-specific staining in flow cytometry, Part I

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? …

March 21, 2013 | PBMC Basics | 1 Comment
Understanding MFI in the context of FACS data

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 …

March 19, 2013 | PBMC Basics | 0 Comments
Natural Killer Cell subtypes and markers in human PBMC

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 …

March 14, 2013 | PBMC Basics | 0 Comments
Generation of CD4+ Th1 cells from human PBMC

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 …

March 5, 2013 | PBMC Basics | 0 Comments
Identification of CD8+ TC1, TC2, and TC17 populations in human PBMC

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 …

February 21, 2013 | PBMC Basics | 0 Comments
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