Subterranean Fishes of the World

3273

2017

AIM 2017

The of study fish immunology by flow and image cytometry is often plagued by a lack of commercially available antibodies or other labeling reagents. Consequently, such work benefits from leveraging intrinsic information from brightfield, light scatter and non-species-specific dyes such as DNA dyes for identifying and enumerating cells types and cell states. However, traditional approached to analysis employing manual gating to identify and score populations from multiparameter data is necessarily error prone and biased by user input, thus confounding efforts to study heterogeneous cell populations such as those present in most primary blood or tissue samples in an unbiased manner. Image cytometry on intrinsic cell features and DNA labeling with t-SNE dimensionality reduction performed in R or FCS Express, or other unsupervised clustering methods, allows identification and study of populations based on multiparameter intrinsic cell features by reducing information from multiparameter data to a number of dimensions more easily interpreted by the analyst, thus aiding in the study of emerging model systems. Here we present data illustrating that identification of several discrete cell populations in the cavefish Astyanax mexicanus hematopoietic tissue can be accomplished using image cytometry and tSNE transformation and that such an approach can be employed to assist the researcher in understanding how experimental manipulations affect multiple cell types within a sample set despite a lack of commercially available antibodies and other reagents.

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