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We describe a method for preparing a cell block from a very sparsely cellular cytology fluid sample. The aim is to ensure that cells are not lost during processing. We developed this method to perform immunohistochemistry on circulating tumour cells (CTC) extracted from the blood of patients with metastatic carcinoma, in which there are typically less than 20 cells in a 10 mL blood sample. The method could also be applicable to other sparsely cellular samples such as cerebrospinal fluid (CSF) samples, or eye vitreous fluid samples.
First, the cells are concentrated into a 90 µL volume of fluid. In the case of CTCs this is obtained from a phosphate buffered saline backwash of a Parsortix CTC blood filter cassette (figure 1A–C; arrows mark CTCs). This filter cassette separates cells based on their size and deformability. Circulating carcinoma cells, and cells from other non-haematological neoplasms, tend to be larger and more rigid than other blood cells. The concentrate could also be from centrifugation of a CSF sample, vitreous humour or similar, with removal of most of the supernatant and …
Handling editor Runjan Chetty.
Contributors The authors agree that they have all made substantial contributions to the conception or design of the work, or the acquisition, analysis or interpretation of data; have been involved in drafting the work or revising it critically for important intellectual content; have/will have final approval of the version published; and agree to be accountable for all aspects of the work in ensuring that questions related to the accuracy or integrity of any part of the work are appropriately investigated and resolved.
Funding This work was funded by ANGLE Europe Ltd (commercial funding), IRAS Project ID: 209687.
Competing interests Taunton and Somerset NHS Foundation Trust employs FGM and JP, and contracts the services of IB. It has received funding from ANGLE Europe Ltd to support this work. ANGLE Europe Ltd employs LS, AH and KMM and is a subsidiary of Angle PLC, the owner of the Parsortix filter technology. The authors have no other competing interests to declare.
Patient consent Not required.
Ethics approval South Birmingham Research Ethics Committee (16/WM/0418).
Provenance and peer review Not commissioned; internally peer reviewed.
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