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Automated REcognition of Tissue-associated Erythrocytes (ARETE)

  1. Top of page
  2. Automated REcognition of Tissue-associated Erythrocytes (ARETE)
  3. Single Cell Network Profiling in Bladder Cancer
  4. T and B lymphocytes become visually distinguishable

Automated microscopic image analysis of immunofluorescence-stained targets on tissue sections is challenged by autofluorescent erythrocytes, which interfere with target segmentation and quantification. To resolve this, the authors developed ARETE, a system for in silico recognition and removal of erythrocytes. To avoid blocking a fluorescence channel, they chose to recognize erythrocytes in the transmission channel only, which also allows adherence to any existing staining protocol. Rather than implement a classical image analysis system by hand, they utilized established machine learning techniques that are used, for example, to recognize smiling faces in state-of-the-art digital cameras. In effect, biologists just had to mark up samples of erythrocytes as well as areas without erythrocytes and the image analysis system was created automatically. By this approach biologists' implicit knowledge - what they simply know but cannot explain - can also be captured. The authors believe that such systems are the future of image analysis. 1

In this issue: page 363

Single Cell Network Profiling in Bladder Cancer

  1. Top of page
  2. Automated REcognition of Tissue-associated Erythrocytes (ARETE)
  3. Single Cell Network Profiling in Bladder Cancer
  4. T and B lymphocytes become visually distinguishable

This study demonstrates the functional characterization of rare cells from bladder cancer (BC) washings as a novel application of multiparametric single cell network profiling (SCNP). Prior studies have reported the prognostic value of applying SCNP to hematologic malignancies, in which live tumor cells are collected in single cell suspensions. Due to standard tissue fixation methodologies, solid tumor cells are typically non-functional upon capture, precluding analytic methods that inform on distinct protein and biochemical pathway alterations. An alternative to conventional solid tumor tissue sampling is the use of fluids containing tumor cells in suspension, such as pleural effusions, bladder washes or peripheral blood (the latter known as circulating tumor cells (CTCs)). This paper describes the quantification of basal and modulated signaling in the PI3K and MAPK pathways, and selective inhibition with a targeted PI3K inhibitor, in bladder washings, providing initial proof of concept for SCNP application to functional analysis of CTCs. 2

In this issue: page 386

T and B lymphocytes become visually distinguishable

  1. Top of page
  2. Automated REcognition of Tissue-associated Erythrocytes (ARETE)
  3. Single Cell Network Profiling in Bladder Cancer
  4. T and B lymphocytes become visually distinguishable

T and B lymphocytes are difficult to distinguish morphologically even with electron microscopy (EM), and antibodies are generally used to make the distinction. In this study, Kono and coworkers investigated the effect of the detergent on the morphology of T and B lymphocytes. FCM and EM observations revealed that B lymphocytes are more likely to lose surface antigens and intracellular organelles than T lymphocytes, which allows the visual distinction between T and B lymphocytes. The ratio of cholesterol to total lipid in T lymphocyte membranes had a tendency to be higher than that in B lymphocyte membranes. The authors demonstrated that cells with differences in cell membrane cholesterol amounts, such as B and T lymphocytes could be identified using an inexpensive detergent, as an alternative to costly antibodies. 3

In this issue, page 396