Method and apparatus for imaging target components in a biological sample using permanent magnets
View Patent ↗A system for enumeration of cells in fluids by image cytometry is described for assessment of target populations such as leukocyte subsets in different bodily fluids or bacterial contamination in environmental samples, food products and bodily fluids. Briefly, all cells in a biological sample are fluorescently labeled, but only the target cells are also magnetically labeled. A small, permanent magnet is inserted directly into the chamber containing the labeled sample. The magnets are coated with PDMS silicone rubber to provide a smooth and even surface which allows imaging on a single focal plane. The cells are illuminated and the images of the fluorescent light emitted by the target cells are captured by a CCD camera. Image analysis performed with a novel algorithm provides a count of the cells on the surface that can be related to the target cell concentration of the original sample.
1. A method for detecting and enumerating a target cell population within a biological specimen said method comprising:
a. obtaining said biological specimen from a subject;
b. labeling said target population with colloidal magnetic nanoparticles coupled to a ligand specific for said cell population within said biological specimen wherein said nanoparticles are BSA coated and approximately 0.2 microns in diameter;
c. isolating said labeled target population wherein said isolating includes art addition of a small PDMS (polydimethylsiloxane)-coated permanent magnet directly in said specimen;
d. removing said magnet from said specimen;
e. acquiring an image of said labeled target population wherein said target population is attached to said magnet along a single focal plane, and
f. analyzing said image to detect and enumerate said labeled target population.
2. The method of claim 1 wherein said target population is CD4 expressing cells.
3. The method of claim 1 wherein said magnet is neodymium.
4. The method of claim 1 wherein said magnet is a disc having a diameter of about 1.6 mm and a height of 0.8 mm.