Trypan Blue: Is It Suitable for Staining of the Endometrium?

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<ul><li><p>Endometrium?</p><p>lated hysterectomy specimens. J Minim Invasive Gynecol. 2013;20:</p><p>667671.</p><p>VisionBlue (trypan blue ophthalmic solution) is a selectiver-</p><p>The use of trypan blue dates back to the 1970s when it was</p><p>in vivo use of trypan blue did not show deleterious effects</p><p>either shadowing or imitating nuclear atypia associatedwith endometrial intraepithelial carcinoma is not a problem2. Awad D, Schrader I, Bartok M, Sudumbrekar N, Mohr A, Gabel D. Bril-1. Tam T, Harkins G, Caldwell T, Zaino R, Hazard D. Endometrial dye</p><p>instillation: a novel approach to histopathologic evaluation of morcel-safety of trypan blue use in surgeries.</p><p>Concern regarding trypan-related morphologic changesI was pleased to read the interesting article by Tam et al[1] published in the Journal of Minimally Invasive Gynecol-ogy. In this prospective pilot case study analysis, the authorsexamined endometrial dye instillation as a novel approachthat may improve the histopathological assessment ofmorcellated specimens.</p><p>Even though their method is useful, I would like tocomment on the authors preference of trypan dye. First,trypan is toxic to living cells [2]. Trypan toxicityrelatedmorphologic changes in surface and glandular epithelialcells were not stressed in the article by Tam et al. Especiallyin an atrophic endometrium, such toxicity-associatedmorphologic changes may either shadow or imitate nuclearatypia associated with endometrial intraepithelial carci-noma because these atypia are usually limited to the sur-face. Second, because trypan specifically stains dead cells[3,4], the living endometrial component might not bestained with dye instillation. Hence, the unstained endome-trial component may be missed from sampling for histo-pathological assessment. Third, the longer exposure ofendometrial tissue to this dye would increase trypantoxicityassociated changes.</p><p>I believe that limiting the use of trypan to in premeno-pausal patients whose endometrial intraepithelial carcinomarisk is relatively lower and avoiding it use in patients inwhom operation time is expected to be longer wouldcontribute to the quality of this method and serve as a futurereference for similar studies.</p><p>Mustafa Ozturk, MDIbrahim Alanbay, MDHaldun Umudum, MD</p><p>Ankara, Turkey</p><p>ReferencesLetters to the Editor</p><p>Trypan Blue: Is It Suitable for Staining of theliant Blue G as protective agent against trypan blue toxicity in human</p><p>retinal pigment epithelial cells in vitro. Graefes Arch Clin Exp Ophthal-</p><p>mol. 2013;251:17351740.</p><p>1553-4650/$ - see front matter 2014 AAGL. All rights reserved.in patients who underwent vitreoretinal surgery [4]. TheAmerican Academy of Ophthalmology has recently reporteda technology assessment on the use of trypan blue in cataractsurgery further suggesting the dyes extensive use and safetyin ophthalmologic surgeries [6]. This further elucidates theused preoperatively to stain the corneal endothelium [3], andis currently used frequently in cataract surgery [4,5]. Thetissue staining agent used as a medical aid in cataract sugery.3. Strober W. Trypan blue exclusion test of cell viability. Curr Protoc Im-</p><p>munol. 2001. May:Appendix 3:Appendix 3B.</p><p>4. Fauque P, Amor AB, Joanne C, Agnani G, Bresson JL, Roux C. Use of</p><p>trypan blue staining to assess the quality of ovarian cryopreservation.</p><p>Fertil Steril. 2007;87:12001207.</p><p>http://dx.doi.org/10.1016/j.jmig.2013.11.003</p><p>Reply to Dr. Ozturk</p><p>We would like to express our appreciation for Dr. Ozturkscomments on our original article [1]. A proper discussion ofthis novel approach evaluating trypan blue instillation forendometrial staining of morcellated hysterectomy specimensis important for patient safety and diagnostic efficiency andaccuracy.</p><p>The first comment regarding trypan being toxic to livingcells is a valid concern. Fortunately, we were able to useVisionBlue (Dutch Ophthalmic USA, Kingston, NH), a ster-ile solution of trypan blue. Each milliliter of VisionBlue con-tains 0.6 mg trypan blue, 1.9 mg sodium monohydrogenorthophosphate (Na2HPO4.2H2O), 0.3 mg sodium dihydro-gen orthophosphate (NaH2PO4*2H2O); 8.2 mg sodiumchloride (NaCl), and water [2]. Trypan blue dye has beenused in numerous surgical settings and in diagnostic evalua-tions to aid in tissue identification or anatomic evaluation.our pathologists encountered. Trypan blue provided pale butdistinct staining of the endometrium on the fresh specimens,which persisted even after overnight fixation. The dye was</p><p>Trypan Blue: Is It Suitable for Staining of the Endometrium?References</p></li></ul>