ADAMⅡ-CD3/CD4/CD8 

ADAMII-CD3/CD4/CD8 Kit is compatible with ADAMII. It is used for counting CD3+, CD3+CD4+, and CD3+CD8+ cell that can be used in research laboratory. 

General Description

ADAMII–CD3/4/8 is a bench-top fluorescence cell counter that counts CD3, CD4, and CD8 expressing cells. ADAMII–CD3/4/8 is an image-based cytometer and it takes up to 75 sets of images from sample loaded in a disposable ADAMII Assay Slide. Each set of images comprises one bright-field image and three fluorescence images. These images are processed to identify CD3 expressing T-lymphocytes, CD4 expressing helper T-lymphocytes, and CD8 expressing cytotoxic T-lymphocytes.

Principle

ADAMII T-cell assay starts with adding an appropriate volume of a sample to a single test tube and mixing the sample with reagent solution containing fluorescence-labeled antibodies. Once the staining step is completed, RBC Lysis Solution is added to lyse red blood cells in the sample. The prepared sample is loaded into a disposable ADAMII Assay Slide and the slide is inserted into ADAMII instrument. ADAMII –CD3/4/8 reports absolute counts of CD3+ T-cells (cells/μL), (2) absolute counts of CD3+/CD4+ T-helper cells (cells/μL), (3) absolute counts of CD3+/CD8+ T-killer cells (cells/μL), (4) ratio of T-helper cells over T-killer cells.

Simple and Easy of use

Flow cytometry is the standard method for enumerating T lymphocyte cells it has some limitations, such as expensive instrumentation and reagent costs, and poor reproducibility between technicians and laboratories. ADAMII is easy to use. Sample preparation steps are shown in the cartoon on the right-hand side. ADAMII doesn’t require routine cleaning and maintenance. Pumping, cleansing and waste bottles are not necessary for the ADAMII.

Results 

The ADAMII simultaneously captures a series of bright field and fluorescent images of the sample in the assay slide and runs a comprehensive image analysis on the images to determine cell counts. With the well-defined geometry of ADAMII Assay Slide, ADAMII reports not only the ratio but also the absolute counts of T cells. Sample images are shown below. 

A) Bright: Total cells are counted. Shapes and sizes of objects are used to eliminate non-cell debris. 

B) CD3 channel: combining with BF channel, CD3+ cells are counted. 

C) CD8 channel: combining with CD3 channel, CD3+/CD8+ cells are counted. 

D) CD4 channel: combining with CD3 channel, CD3+/CD4+ cells are counted.

Accuracy

Total 30 whole blood samples were used to compare ADAMII and flow cytometry. Linear regression analysis revealed a close correlation between the data for the absolute number of human helper/inducer, suppressor/cytotoxic, and total T lymphocytes obtained with the ADAMII and flow cytometry (Absolute number of CD3+, R2 = 0.998; Absolute number of CD3+/CD4+, R2 = 0.993; Absolute number of CD3+/CD8+, R2=0.992). Detailed data are shown in the graphs below.  

Precision (within-run)

Total T lymphocytes (CD3+), helper/inducer(CD3+CD4+), suppressor/cytotoxic(CD3+CD8+) of precision test was conducted using whole blood with 6 different concentrations. Each concentration was tested in 6 biological replicates and 3 technical replicates per preparation. The coefficient of variation of total T lymphocytes (CD3+) and T lymphocytes subsets (CD3+/CD4+, and CD3+/CD8+) were below 10%. Detailed data are shown in the tables below. 

Operator to operator

Three operators performed CD3/CD4/CD8 count at low concentration and showed low CV values which demonstrated high accuracy in operator to the operator comparison test. 

Instrument to instrument

Three instruments performed CD3/CD4/CD8 count at low concentration and showed low CV values which demonstrated high accuracy in instrument to the instrument comparison test. 

Specifications