Name: Tubal Cytology of the Fallopian Tube as a Promising Tool for Ovarian Cancer Early Detection (Video) | JoVE Journal
Uploaded: 2017-07-25T13:00:00.000+00:00
Duration: 8 min 9 s
Description: Learn tubal cytology for early ovarian cancer detection using direct fallopian tube sampling. This method identifies malignant cytologic features to improve diagnostic accuracy in gynecologic oncology.

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5 minutes

The authors would like to acknowledge the Department of Pathology, University of Arizona, for the financial support. The project is partially supported by the Mark and Jane Gibson endowment fund to WZ.

An Institutional Review Board approval was received from University of Arizona for conducting this study. Informed patient consent forms were obtained.
1. Patient Selection
NOTE: An Institutional Review Board approval was obtained from University of Arizona. A total of 38 patients were recruited. The patients&#39; ages ranged from 32 to 86 years old with a mean of 55 years, of whom 26 patients were post-menopausal and 12 patients were pre-menopausal.
<ol>
	<li>Obtain informed consent from each patient.</li>
</ol>
2. Fallopian Tube Cells Collection Procedure
NOTE: The patients included in the current study were all scheduled for total hysterectomy and bilateral salpingo-oophorectomy for either prophylactic risk reduction or malignancy. The details of surgery were unknown for pathologist who performed the cytology study.
<ol>
	<li>Immediately after surgical excision, examine the fresh tissue specimen including uterus, bilateral ovaries, and bilateral fallopian tubes carefully to identify the bilateral fallopian tube.</li>
	<li>Collect and place fallopian tube cells into the preservative solution (see Table of Materials) vial within 30 min of clamping the blood supply; the recommended preservative is a methanol-based, buffered preservative solution. 
	NOTE: Use a gentle touch brush for the tubal cell collection (upcoming steps). With the exception of rare cases in which the fallopian tube is submitted detached from the uterus, inserting a brush head to collect cells from the tubal fimbria occurs typically when the fallopian tube is attached to the uterus.
	<ol>
		<li>With the help of a small forceps, insert the brush (see Table of Materials) into the lumen of the fallopian tube through the fimbriae end. Slowly rotate clockwise and move the brush forward through the infundibular portion of the fallopian tube, then the ampulla until it reaches the isthmus.</li>
		<li>Slowly rotate and reverse the brush counter-clockwise to pull out the brush. Ensure that the maximal speed of the brush within the lumen is slower than 1 cm/s.</li>
		<li>Rinse the brush in the preservative solution (in a vial) by rotating and swirling the brush 10 times in the solution.</li>
		<li>Tighten the vial cap.</li>
		<li>Record the patient&#39;s information and the laterality of the specimen.</li>
		<li>Store the vial in a 4 &#176;C refrigerator (for up to) 6 months.</li>
	</ol>
	</li>
</ol>
3. Tubal Cytology Slide Preparation
<ol>
	<li>Load the sample vial into the automated processor for slide preparation. 
	NOTE: We use the automated processor for slide preparation. The following steps are performed automatically after the sample vial is placed into the processor: (i) The cells and debris are separated and dispersed through the rotation of the test filter within the sample vial. (ii) Tubal cells are collected from the exterior surface of the filter by a gentle vacuum. (iii) The filter is inverted and pressed against the slide to let the cells to adhere to the circular area within the slide. (iv) The slide is further fixed in a cell fixative bath and ready for staining and cytology analysis. Unfortunately, there is no alternative manual method, which can yield results with a similar quality.</li>
</ol>
4. Modified Papanicolaou Staining Protocol
<ol>
	<li>Stain the slides made from step 3.1 by running the automatized non-gyn stain procedure, as illustrated in Table 1. 
	NOTE: The staining method represents a modification of the Papanicolaou staining technique that we use routinely for non-gynecologic specimens. We use an automated tissue processor to stain some slides. We chose the Non-Gyn staining procedure over the Gyn staining procedure in this study. The Eosin used is EA-65, instead of the EA-50 used for gynecological specimens (Pap smear specimen). In addition, OG-6 in the Gyn staining procedure, which is mainly for squamous cell staining, is not included in the staining procedure since no squamous cells are expected to be seen in this study. EA65 (Eosin Azure) is a counter stain solution comprised of 3 dyes: Eosin Y, Fast green FCF, and Bismarck brown Y. The procedure is illustrated in Table 1.</li>
	<li>Perform a manual quick Pap stain procedure as follows.
	<ol>
		<li>Dip the slide made in step 3.1 into 95% ethanol, 10 times, with each dip taking about 1 s.</li>
		<li>Dip the slide into H2O, 10 times, with each dip taking about 1 s.</li>
		<li>Emerge the slide in hematoxylin for 10 - 60 s.</li>
		<li>Dip the slide into H2O, 10 times, with each dip taking about 1 s.</li>
		<li>Dip the slide into 95% ethanol, 10 times, with each dip taking about 1 s.</li>
		<li>Emerge the slide in EA65 for 1 - 3 min.</li>
		<li>Dip the slide into H2O, 10 times, with each dip taking about 1 s.</li>
		<li>Dip the slide into 95% ethanol, 10 times, with each dip taking about 1 s.</li>
		<li>Dip the slide in xylene until the slide becomes clear. 
		NOTE: This is a modified Papanicolaou staining procedure. This method is used as a more rapid stain method for cases with limited time or space like on-site fine needle aspiration and STAT specimens. This procedure provides comparable quality staining as the automatized Non-Gyn stain procedure. The procedure is illustrated in Table 2.</li>
	</ol>
	</li>
</ol>
5. Spin Cells onto Slides
NOTE: Three cytospin slides are made for each specimen. One slide is H&#38;E stained for the morphological comparison with the cytology slide. Two unstained slides were made for the future IHC staining study. The detailed steps of this section are not the focus of this paper; therefore, we will not discuss them in extended details.
<ol>
	<li>Prepare a cytocentrifuge with a labeled slide, specimen funnel and filter for each sample to be examined.</li>
	<li>Concentrate the cells by centrifuging for 5 min at 200 x g.</li>
	<li>Remove about half of the supernatant and then re-suspend the cells by gentle pipetting.</li>
	<li>Add 200 &#181;L of each cell suspension to a specimen funnel.</li>
	<li>Spin at 250 x g for 5 min. Carefully remove the slide(s) from the cytocentrifuge and transfer them to 95% Ethanol for fixation.</li>
	<li>Air dry prior to staining and for long term storage. 
	NOTE: Depending on the cell density on the final slides, step 5.2 and step 5.3 can be skipped or adjusted. In this study, we require at least 2 cell clusters present per high power view for adequate cellular density.</li>
</ol>
6. Sectioning and Extensively Examining the Fimbriated End (SEE-FIM) Protocol
<ol>
	<li>Fix the entire specimen (including uterus, bilateral fallopian tubes, and ovaries) in 10% formalin prior to grossing to minimize exfoliation.</li>
	<li>Gently and carefully detach the fallopian tubes from the uterus at the isthmus using small forceps and a scissors. If adhesion is present, carefully dissect the fimbriated end from the ovary surface using small forceps and a scissors; these steps are crucial to minimize tumor cross contamination between ovary and fallopian tube due to their intimate proximity.</li>
	<li>Amputate the infundibulum and fimbriae segment (the distal 1.0 - 2.0 cm of fallopian tube) from the rest of the specimen.</li>
	<li>Section the infundibulum and fimbriae longitudinally at 2 mm intervals. Submit all sections in toto for histologic examination.</li>
	<li>Section the isthmus and ampulla at 2 - 3 mm intervals and submit entirely.</li>
</ol>

<ol><li>Carlson, J. W., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Serous+tubal+intraepithelial+carcinoma%3A+its+potential+role+in+primary+peritoneal+serous+carcinoma+and+serous+cancer+prevention%5BTitle%5D)+AND+%22J+Clin+Oncol%22%5BJournal%5D)">Serous tubal intraepithelial carcinoma: its potential role in primary peritoneal serous carcinoma and serous cancer prevention.</a> J Clin Oncol. 26 (25), 4160-4165 (2008).</li>
<li>Kurman, R. J., Shih Ie, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+origin+and+pathogenesis+of+epithelial+ovarian+cancer%3A+a+proposed+unifying+theory%5BTitle%5D)+AND+%22Am+J+Surg+Pathol%22%5BJournal%5D)">The origin and pathogenesis of epithelial ovarian cancer: a proposed unifying theory.</a> Am J Surg Pathol. 34 (3), 433-443 (2010).</li>
<li>Lee, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((A+candidate+precursor+to+serous+carcinoma+that+originates+in+the+distal+fallopian+tube%5BTitle%5D)+AND+%22J+Pathol%22%5BJournal%5D)">A candidate precursor to serous carcinoma that originates in the distal fallopian tube.</a> J Pathol. 211 (1), 26-35 (2007).</li>
<li>Li, J., Fadare, O., Xiang, L., Kong, B., Zheng, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Ovarian+serous+carcinoma%3A+recent+concepts+on+its+origin+and+carcinogenesis%5BTitle%5D)+AND+%22J+Hematol+Oncol%22%5BJournal%5D)">Ovarian serous carcinoma: recent concepts on its origin and carcinogenesis.</a> J Hematol Oncol. 5, 8(2012).</li>
<li>Przybycin, C. G., Kurman, R. J., Ronnett, B. M., Shih Ie, M., Vang, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Are+all+pelvic+%28nonuterine%29+serous+carcinomas+of+tubal+origin%3F%5BTitle%5D)+AND+%22Am+J+Surg+Pathol%22%5BJournal%5D)">Are all pelvic (nonuterine) serous carcinomas of tubal origin?</a> Am J Surg Pathol. 34 (10), 1407-1416 (2010).</li>
<li>Zheng, W., Fadare, O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Fallopian+tube+as+main+source+for+ovarian+and+pelvic+%28non-endometrial%29+serous+carcinomas%5BTitle%5D)+AND+%22Int+J+Clin+Exp+Pathol%22%5BJournal%5D)">Fallopian tube as main source for ovarian and pelvic (non-endometrial) serous carcinomas.</a> Int J Clin Exp Pathol. 5 (3), 182-186 (2012).</li>
<li>Carcangiu, M. L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Atypical+epithelial+proliferation+in+fallopian+tubes+in+prophylactic+salpingo-oophorectomy+specimens+from+BRCA1+and+BRCA2+germline+mutation+carriers%5BTitle%5D)+AND+%22Int+J+Gynecol+Pathol%22%5BJournal%5D)">Atypical epithelial proliferation in fallopian tubes in prophylactic salpingo-oophorectomy specimens from BRCA1 and BRCA2 germline mutation carriers.</a> Int J Gynecol Pathol. 23 (1), 35-40 (2004).</li>
<li>Leunen, K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Prophylactic+salpingo-oophorectomy+in+51+women+with+familial+breast-ovarian+cancer%3A+importance+of+fallopian+tube+dysplasia%5BTitle%5D)+AND+%22Int+J+Gynecol+Cancer%22%5BJournal%5D)">Prophylactic salpingo-oophorectomy in 51 women with familial breast-ovarian cancer: importance of fallopian tube dysplasia.</a> Int J Gynecol Cancer. 16 (1), 183-188 (2006).</li>
<li>Piek, J. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Dysplastic+changes+in+prophylactically+removed+Fallopian+tubes+of+women+predisposed+to+developing+ovarian+cancer%5BTitle%5D)+AND+%22J+Pathol%22%5BJournal%5D)">Dysplastic changes in prophylactically removed Fallopian tubes of women predisposed to developing ovarian cancer.</a> J Pathol. 195 (4), 451-456 (2001).</li>
<li>Crum, C. P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+distal+fallopian+tube%3A+a+new+model+for+pelvic+serous+carcinogenesis%5BTitle%5D)+AND+%22Curr+Opin+Obstet+Gynecol%22%5BJournal%5D)">The distal fallopian tube: a new model for pelvic serous carcinogenesis.</a> Curr Opin Obstet Gynecol. 19 (1), 3-9 (2007).</li>
<li>Jarboe, E. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Tubal+and+ovarian+pathways+to+pelvic+epithelial+cancer%3A+a+pathological+perspective%5BTitle%5D)+AND+%22Histopathology%22%5BJournal%5D)">Tubal and ovarian pathways to pelvic epithelial cancer: a pathological perspective.</a> Histopathology. 53 (2), 127-138 (2008).</li>
<li>Menon, U. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Ovarian+cancer+screening+has+no+effect+on+disease-specific+mortality%5BTitle%5D)+AND+%22Evid+Based+Med%22%5BJournal%5D)">Ovarian cancer screening has no effect on disease-specific mortality.</a> Evid Based Med. 17 (2), 47-48 (2012).</li>
<li>Buys, S. S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Effect+of+screening+on+ovarian+cancer+mortality%3A+the+Prostate%2C+Lung%2C+Colorectal+and+Ovarian+%28PLCO%29+Cancer+Screening+Randomized+Controlled+Trial%5BTitle%5D)+AND+%22JAMA%22%5BJournal%5D)">Effect of screening on ovarian cancer mortality: the Prostate, Lung, Colorectal and Ovarian (PLCO) Cancer Screening Randomized Controlled Trial.</a> JAMA. 305 (22), 2295-2303 (2011).</li>
<li>Otsuka, I., Kameda, S., Hoshi, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Early+detection+of+ovarian+and+fallopian+tube+cancer+by+examination+of+cytological+samples+from+the+endometrial+cavity%5BTitle%5D)+AND+%22Br+J+Cancer%22%5BJournal%5D)">Early detection of ovarian and fallopian tube cancer by examination of cytological samples from the endometrial cavity.</a> Br J Cancer. 109 (3), 603-609 (2013).</li>
<li>Dhanani, M., Nassar, A., Dinh, T. <a target="_blank" href="http://www.jascyto.org/article/S2213-2945(14)00293-2/abstract">Classification of Fallopian Tube Cytology Sampling to Develop a Model for Future Peritoneal and Ovarian Cancer Screening.</a> J Am Soc Cytopathol. 3 (5), S35-S36 (2014).</li>
<li>Rodriguez, E. F., Lum, D., Guido, R., Austin, R. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Cytologic+findings+in+experimental+in+vivo+fallopian+tube+brush+specimens%5BTitle%5D)+AND+%22Acta+Cytol%22%5BJournal%5D)">Cytologic findings in experimental in vivo fallopian tube brush specimens.</a> Acta Cytol. 57 (6), 611-618 (2013).</li>
<li>Chen, H., Klein, R., Arnold, S., Chambers, S., Zheng, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Cytologic+studies+of+the+fallopian+tube+in+patients+undergoing+salpingo-oophorectomy%5BTitle%5D)+AND+%22Cancer+Cell+Int%22%5BJournal%5D)">Cytologic studies of the fallopian tube in patients undergoing salpingo-oophorectomy.</a> Cancer Cell Int. 16, 78(2016).</li>
</ol>

The authors declare that they have no competing financial interests.

Prophylactic bilateral salpingectomy or salpingo-oophorectomy has been shown to decrease the risk of developing HGSC in high risk populations, such as women with BRCA gene mutations and strong familial cancer history. However, the sterility and surgical menopause caused by the surgery are serious consequences and make for a difficult decision, especially for women of childbearing age. The previous study has shown excellent correlation between tubal cytology and histology17. We believe that tubal c...

Ovarian high-grade serous carcinoma (HGSC) is the most common and lethal type of ovarian cancer, and remains a major threat to the public health. In the past decade, researchers have suggested that the vast majority of HGSC cases arise from the fallopian tube instead of the ovary itself 1,2,3,4,5,6. Serous tubal intraepithelial carcinoma (STIC) is widely accepted as the precursor of HGSC 1,7,8,9,10,11. So far, early detection of ovarian cancer remains difficult. The current screening protocol with combined cancer antigen 125 (CA-125) and transvaginal ultrasound has shown little effect on patient mortality12,13. Endometrial sampling for ovarian cancer detection has shown an extremely low sensitivity14. Two pioneer studies15,16 explored the usage of laparoscopic direct sampling of benign fallopian tubes and characterized the cytological features of benign tubal epithelium. We believe that direct sampling from the fallopian tube can also be a practical and straightforward way to detecting STIC or HGSC at an early stage when the tumor is not visualized by imaging.
Although the ultimate goal is the utility of minimally invasive laparoscopic screening in patients with high-risk factors, the immediate aims of current study are: 1) To establish the baseline cytological features of benign tubal epithelia; and 2) To test the sensitivity and specificity of tubal cytology in detecting ovarian cancers and cancer precursors. We therefore developed the following baseline tubal cytology method to test the effectiveness of this protocol in detection of HGSC and its precursor STIC17. In this study, we took advantage of the large volume of regularly received surgical specimens, and performed direct sampling of the surgically excised fallopian tube in addition to the routine grossing procedures.
Our recent study17 showed that the cytological diagnosis of malignant or suspicion for malignancy is highly correlated with the histological diagnosis of HGSC (100%). In addition, the tubal cytological evaluation identified intratubal neoplasia in the only two histologically confirmed STIC cases involved in this study. We believe that tubal cytology has great potential in early detection and will provide valuable information for patient management.

<table><tbody><tr><td>Cytobrush plus GT gentle touch of Medscand sample collection kit</td><td>CooperSurgical</td><td>C0112</td><td></td></tr><tr><td>ThinPrep preservCyt solution</td><td>HOLOGIC</td><td>234005</td><td></td></tr><tr><td>ThinPrep 2000 Processor</td><td>HOLOGIC</td><td>70031-001</td><td></td></tr><tr><td>Papanicolaou Stain, EA 65</td><td>sigma aldrich</td><td>HT40432</td><td></td></tr><tr><td>95% Ethanol</td><td>sigma aldrich</td><td>652261</td><td></td></tr><tr><td>100% Ethanol</td><td>sigma aldrich</td><td>493538</td><td></td></tr><tr><td>Xylene</td><td>sigma aldrich</td><td>214736</td><td></td></tr><tr><td>Hematoxylin</td><td>sigma aldrich</td><td>H9627</td><td></td></tr><tr><td>Sakura Tissue Tek 2000 tissue processor</td><td>SAKURA</td><td>TISSUE-TEK VIP 2000</td><td></td></tr><tr><td>Mayo Dissecting Scissors,5.5 straight</td><td>Pilgrim medical equiment</td><td>FA710-45</td><td></td></tr><tr><td>Chemware Forceps</td><td>United state plastic corp</td><td>76122</td><td></td></tr><tr><td>Tissue-Tek Accu-Edge Trimming blades,short</td><td>SAKURA</td><td>4785</td><td></td></tr><tr><td>Tissue-Tek Accu-Edge Trimming blades handle,short</td><td>SAKURA</td><td>4786</td><td></td></tr></tbody></table>

tubal cytology of the fallopian tube as a promising tool for ovarian cancer early detection

Tubal Cytology of the Fallopian Tube as a Promising Tool for Ovarian Cancer Early Detection

Currently, it is widely accepted that the vast majority of ovarian high-grade serous carcinoma (HGSC) originate from the fallopian tube. However, due to the lack of markers or tools for the ovarian cancer identification, the early detection of HGSC remains challenging. Direct sampling of the fallopian tube can enhance sensitivity for detection of neoplastic cells when the tumor is not grossly visible. We developed a procedure to collect fallopian tube cells directly from freshly received surgical specimens, which has shown excellent correlation with histological findings. This approach lays a foundation for the future utility of minimally invasive laparoscopic screening in high-risk patient populations.

Our previous study showed that sampling directly from the excised fallopian tube using a gentle touch brush could yield quantitatively and qualitatively adequate specimen for further cytological evaluation. In addition, the combination of automated slide preparation and modified Papanicolaou staining enable the pathologist to better visualize the cytological details to make an accurate diagnosis. An example of a manual quick pap stain of a specimen from a benign fallopian tube is illustra...

We explored a tubal cytologic method by sampling the fallopian tube directly post-surgical excision as a tool of ovarian cancer early detection. Here, we present a protocol to collect fallopian tube cells from freshly received surgical specimens.

Currently, it is widely accepted that the vast majority of ovarian high-grade serous carcinoma (HGSC) originate from the fallopian tube. However, due to ...

The overall goal of this protocol is to direct the sample fallopian tube epithelium from the salpingo oophorectomy specimen. For the identification of tubal cytologic features that allow the differentiation of benign tissue from malignant or pre-malignant conditions. This cytology method will help answer some key questions in the gynecologic oncology field regarding early detection of ovarian and fallopian tube cancer.The advantage of this technique is it requires direct sampling of the fallopian tube cells. Another advantage of this technique is that it is a direct and simple method. In the future, it maybe applied during laparoscopic screening for ovarian or fallopian tube cancers in women who are high risk for those diseases.A visual demonstration of the collection method is important as the collection steps can be difficult to learn. We want to make sure that there is minimal disturbance to the fallopian tube epithelium as the histologic diagnosis of the fallopian tube is important in the end, which will then impact clinical management. Demonstrating this procedure is Dr.Howard Chen, whose currently our fourth year pathology resident at the University of Arizona.Immediately after obtaining the tissue, confirm that the uterus, bilateral fallopian tubes and bilateral ovarian are intact. Using small forceps, carefully insert the cytobrush into the lumen of the fallopian tube through the end and slowly rotate the brush clockwise. Moving the brush into the tube, through the infundibular and ampulla regions.When the brush reaches the isthmus, slowly begin rotating the brush in the counter-clockwise direction while pulling the brush back out of the fallopian tube. Next, rotate and swirl the fallopian cell coated brush in freshly prepared preservative solution 10 times. Then tightly cap the sample vial and label it with the patient's information and the laterality of the specimen.The cells can then be stored at four degree celsius for up to six months. Before staining the cells, load the sample vial into an automated processor for slide preparation. When the slides are ready, dip the samples into 95%ethanol, 10 times for one second per dip.Followed by 10 dips in water for one second per dip. Next, submerge the slides in hematoxylin for 10 to 60 seconds, followed by a second water ethanol wash. After the last ethanol immersion, submerge the slides in eosin azure for one to three minutes.Followed by another water ethanol rinse. Then dip the slides in xylene, until the samples become clear. To obtain cyto spins of the harvested fallopian tube cells, first label three slides for each cell sample with the appropriate patient information.Next, attach a filter and a specimen funnel to each slide, and pellet the fallopian tube cells by bench top centrifugation. Remove about half of the supernatant and re-suspend the cells with gentle pipetting, then load 200 microliters of the single cell suspension into each specimen funnel and load the slide assemblies into the cytocentrifuge. At the end of the centrifugation, carefully detach the slides from the funnels and filters and fix the cytospun cells at 95%ethanol for 30 seconds.For sectioning and analysis of the fimbriated end of the fallopian tube sample, first fix the entire specimen for two to four hours in 10%formalin. Next, use small forceps and scissors to carefully detach the fallopian tubes from the uterus at the isthmus, if an adhesion is present, use the forceps and scissors to carefully dissect the fimbriated end from the ovary surface. Amputate the infundibulum and fimbria segment at the distal one to two centimeter end of the fallopian tube and section the infundibulum and fimbria, longitudinally at two millimeter intervals.Submit all of the infundibulum and fimbria sections in total for histologic examination. Then section the isthmus and ampulli, three to five millimeter intervals, collectively submitting the specimens in their entirety. The brush rotation maneuver results in an adequate sampling of the specimen and the manual quick PAP stain provides a good resolution of the nuclear details and the cytoplasmic transparency.Both of which are essential for an accurate evaluation of the tubule cells. Although similar results can be obtained by cytocentrifugation, the quick PAP stain procedure results in an ultimately cleaner background and sharper resolution between the nuclear details in the cytoplasmic transparency. The arrow pointing to the ciliates of the ciliated epithelium.Histologic analysis of the fimbria segment sections can be used to access the morphology of the small ciliated and large non-ciliated secretary cell populations. In the ampulla region of the same section, finger-like fimbria can be observed, as well as a mild disturbance in the villi structure. The cytological appearance of malignancy can be determined by the presence of three dimensional clusters composed of cells with prominent nucleoli and anisonucleosis.Hygrade serious carcinoma-like cell groups can also be observed, as shown within the circle. Within the angulated sheets of normal epithelia cells, as pointed by arrows. Once mastered, this collection and staining procedure can be completed within two hours if performed properly.While attempting this procedure, it is important to avoid cross-contamination with other tissues which can then confound the results. Following this procedure, molecular analysis or immunohistochemistry can be performed on the fake cells, if the cytologic findings are equivocal. This technique lays a foundation for future application to laparoscopic or minimally invasive surgery for screening of women at high risk for ovarian and fallopian tube cancer.After watching this video, you will have a good understanding of how to collect and process tubule epithelium from resected salpingo-oophorectomy specimens.

tubal-cytology-fallopian-tube-as-promising-tool-for-ovarian-cancer

Research

JoVE Journal

Medicine

The fallopian tube (FT) is emerging as an alternative site of origin for serous ovarian carcinoma (SOC).  This protocol describes a novel method for the isolation and ex vivo culture of fallopian tube epithelial cells.  This system recapitulates the in vivo epithelium and allows the study of SOC pathogenesis.

Ex Vivo Culture of Primary Human Fallopian Tube Epithelial Cells

We describe methodologies for establishing in vitro heterotypic three-dimensional models comprising ovarian fibroblasts and normal ovarian surface or ovarian cancer epithelial cells. We discuss the use of these models to study stromal-epithelial interactions that occur during ovarian cancer development.

Heterotypic Three-dimensional In Vitro Modeling of Stromal-Epithelial Interactions During Ovarian Cancer Initiation and Progression

In this video, we demonstrate a modified protocol of Papanicolaou staining to study tubal cytology. This technique can be used to visualize the cytological features of malignant and non-malignant fallopian tube epithelial cells.

Papanicolaou Staining: A Technique to Visualize the Cytological Features of Tubal Cells

We describe a non-invasive animal imaging platform that allows the detection, quantification, and monitoring of ovarian cancer growth and recurrence. This intra-peritoneal xenograft model mimics the clinical profile of patients with ovarian cancer.

Murine Model for Non-invasive Imaging to Detect and Monitor Ovarian Cancer Recurrence

We describe an ovarian fad pad transplantation assay suitable for studies of normal and transformed epithelia of the female reproductive tract. The mouse fat pad allows transplantation of large tissue fragments, is easily accessible for surgery and imaging, and provides the most favorable native environment for tissues of Müllerian origin.

Transplantation Into the Mouse Ovarian Fat Pad

A protocol is presented to detect circulating ovarian tumor cells utilizing a custom-made photoacoustic flow system and targeted folic acid-capped copper sulfide nanoparticles.

Ovarian Cancer Detection Using Photoacoustic Flow Cytometry

This protocol presents techniques to laparoscopically excise ovarian endometrioma, to perform adhesiolysis with sparing electrosurgical application, and to employ intraoperative chromopertubation to assess for genital tract patency. This systematic approach will facilitate optimal endometriosis management, guide concomitant adnexal surgeries, and enhance post-surgical fertility outcomes.

Surgical Techniques to Optimize Ovarian Reserve during Laparoscopic Cystectomy for Ovarian Endometrioma

This protocol describes the identification and resection of sentinel lymph nodes to make the operation as easy and minimally invasive as possible.

Sentinel Lymph Node Mapping and Biopsy for Endometrial Cancer at Early Stage with Laparoscopy

We report a coregistered ultrasound and photoacoustic imaging protocol for the transvaginal imaging of ovarian/adnexal lesions. The protocol may be valuable to other translational photoacoustic imaging studies, especially those using commercial ultrasound arrays for the detection of photoacoustic signals and standard delay-and-sum beamforming algorithms for imaging.

A Coregistered Ultrasound and Photoacoustic Imaging Protocol for the Transvaginal Imaging of Ovarian Lesions

This protocol describes a one-day method for the isolation of human fallopian tube epithelial cells. Isolated epithelial cells can be plated in 2-dimensional (2D) culture or dissociated into single-cell suspensions and utilized in downstream experiments, including flow cytometry and single-cell RNA sequencing.

An Efficient Method for Extracting Human Fallopian Tube Epithelia for Single-cell Analyses

This protocol details the development of high-grade serous ovarian carcinoma (HGSOC) mouse models in vitro and in vivo. From the derivation of murine fallopian tube organoids, their genetic modification to recapitulate human HGSOC genetics, and their introduction in the ovarian bursa of syngeneic mice for in vivo development of tumors.

Development of Mouse-Derived Organoid Lines from Fallopian Tube Epithelial Cells for High Grade Serous Ovarian Carcinoma Modeling

Learn tubal cytology for early ovarian cancer detection using direct fallopian tube sampling. This method identifies malignant cytologic features to improve diagnostic accuracy in gynecologic oncology.

Tubal Cytology of the Fallopian Tube as a Promising Tool for Ovarian Cancer Early Detection

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