Liao, HaihuiSheridan, ToddCosar, EdizOwens, ChristopherZuo, TaoWang, XiaofeiAkalin, AliKandil, Dina HDresser, KarenFogarty, KevinBellve, KarlBaer, Christina EFischer, Andrew2023-07-132023-07-132022-02-11Liao H, Sheridan T, Cosar E, Owens C, Zuo T, Wang X, Akalin A, Kandil D, Dresser K, Fogarty K, Bellve K, Baer C, Fischer A. Deconvolution microscopy: A platform for rapid on-site evaluation of fine needle aspiration specimens that enables recovery of the sample. Cytopathology. 2022 May;33(3):312-320. doi: 10.1111/cyt.13106. Epub 2022 Feb 11. PMID: 35102620; PMCID: PMC9305921.1365-230310.1111/cyt.1310635102620https://hdl.handle.net/20.500.14038/52276Context: Rapid on-site evaluation (ROSE) optimises the performance of cytology, but requires skilled handling, and smearing can make the material unavailable for some ancillary tests. There is a need to facilitate ROSE without sacrificing part of the sample. Objective: We evaluated the image quality of inexpensive deconvolution fluorescence microscopy for optically sectioning non-smeared fine needle aspiration (FNA) tissue fragments. Design: A portion of residual material from 14 FNA samples was stained for 3 min in Hoechst 33342 and Sypro™ Red to label DNA and protein respectively, transferred to an imaging chamber, and imaged at 200× or 400× magnification at 1 micron intervals using a GE DeltaVision inverted fluorescence microscope. A deconvolution algorithm was applied to remove out-of-plane signal, and the resulting images were inverted and pseudocoloured to resemble H&E sections. Five cytopathologists blindly diagnosed 2 to 4 representative image stacks per case (total 70 evaluations), and later compared them to conventional epifluorescent images. Results: Accurate definitive diagnoses were rendered in 45 (64%) of 70 total evaluations; equivocal diagnoses (atypical or suspicious) were made in 21 (30%) of the 70. There were two false positive and two false negative "definite" diagnoses in three cases (4/70; 6%). Cytopathologists preferred deconvolved images compared to raw images (P < 0.01). The imaged fragments were recovered and prepared into a ThinPrep or cell block without discernible alteration. Conclusions: Deconvolution improves image quality of FNA fragments compared to epifluorescence, often allowing definitive diagnosis while enabling the ROSE material to be subsequently triaged.enhttp://creativecommons.org/licenses/by-nc-nd/4.0/deconvolutionfine-needle aspirationfluorescence microscopyoptical sectioningrapid on-site evaluationJournal ArticleCytopathology : official journal of the British Society for Clinical Cytology