Air-dried Smears Research Articles

Characteristics of chronic lymphocytic leukemia in Japan: Comprehensive analysis of the CLLRSG-01 study.

Chronic lymphocytic leukemia (CLL) is rare in Japan. We conducted the nationwide, prospective observational study CLLRSG-01 to clarify the current state of CLL in Japan and to make accurate international comparisons by preparing naturally air-dried smears like those used in other countries. Of the 201 untreated patients enrolled and evaluated, 119 were diagnosed with CLL and 82 with non-CLL mature B-cell neoplasms, based on the WHO classification. Of the 119 CLL patients, 90 were classified as typical and 29 as atypical according to FAB classification morphology, with the proportion of atypical CLL consistent with reports from other countries. Immunophenotypic analyses by flow cytometry showed that 55% of Japanese CLL patients had a Matutes score of 4 or higher, which is lower than the rate of about 90% in Europeans. Mutated IGHV was identified in 80% of Japanese CLL patients, which is a higher rate than in Western patients. The most frequent IGHV gene was VH3-30 (15%), followed by VH3-23 (12%) and VH4-34 (10%). VH1-69, the most common gene in Western countries, was identified in only one patient. These results indicate that the pattern of immunophenotypes and IGHV gene usage in Japanese CLL patients differs from that in Western patients.

Role of FNAC in Diagnosis of Malignant Lymphadenopathy: A Five-Year Cross-sectional Study at A Tertiary Care Centre, Mumbai, India

Introduction: Fine Needle Aspiration Cytology (FNAC) of the lymph node is a simple tool in the diagnosis of suspected and unsuspected primary and secondary lymph node malignancy. It is a useful first-line investigation to distinguish between infective and malignant lymphadenopathy, especially in a low-resource setting like our country. Aim: To study the utility of FNAC in the diagnosis of lymph node malignancy and to evaluate the efficacy of cytology in diagnosing the primary site of malignancy in case of metastasis. Materials and Methods: A cross-sectional retrospective and prospective study was conducted for a five-year period from July 2015 to June 2020, in the pathology department of Dr. RN Cooper Municipal General Hospital, Mumbai, Maharashtra, India. A total of 112 patients diagnosed with malignancy on FNAC of lymph nodes were studied. The FNAC procedure was performed by cytopathologists using a 23/24G needle attached to a 10 ml syringe. The alcohol-fixed smears were stained with Haematoxylin & Eosin (H&E) stain and Papanicolaou (PAP) stains, while the airdried smears were stained with Giemsa stain. Diagnosis was based on cytomorphological features and clinical presentation. Results: A total of 112 patients were diagnosed with malignancy on FNAC, of which 90 were males and 22 were females, with a male-to-female ratio of 4:1. Cytological analysis of these nodes revealed metastasis in 98 cases (87.50%) and lymphoma in 14 cases (12.50%). The maximum number of patients were in the 51-60 years age group, with 38 cases (33.92%). The most common site of involvement was cervical lymph nodes, with 75 cases (66.96%). The most common cytological diagnosis was metastatic squamous cell carcinoma in 65 cases (58.02%), followed by metastatic un-differentiated carcinoma in 26 cases (23.20%). Conclusion: Fine Needle Aspiration (FNA) is a simple, safe, accurate, cost-effective, and valuable tool in the evaluation of malignant lymphadenopathy. Malignant diseases were easily diagnosed by this simple diagnostic procedure. In the case of a diagnosis of lymphoma, surgery can be completely avoided. It helps in planning further surgical management for metastatic disease, where definitive operative intervention can be performed in one session. In the case of an un-detected primary tumour, FNAC directs further investigations towards the possible primary site.

Environment Friendly Rehydrants for Air-Dried Oral Buccal Smears - A Comparative Study with Conventional Method.

Cytopathology is defined as a branch of pathology which involves studying diseases and their diagnosis. The conventional method involves fixing the cells in alcohol, but its poor availability in ill-equipped areas can pose problems. One solution to this problem is air-drying of smears followed by rehydration. In this study, we intend to assess the cytomorphologic changes in air-dried smears rehydrated using various agents and to correlate them with the smears prepared using conventional wet technique. Also, we attempt to find the alternative to alcohol which can be used in less equipped areas. Three smears each were prepared from 24 patients (12 healthy and 12 with habits). A total of 72 smears were prepared. One smear from each patient was used as control group (alcohol fixation - total number: 24 smears). The remaining two smears were air-dried and rehydrated and were taken as the test group (test group A - rehydrated with aq. glycerine; total number: 24 smears, test group B - rehydrated with coconut oil; total number: 24 smears) for 5-10 minutes. Then, all 72 slides were stained by standard Papanicolaou stain. The slides were examined for cytomorphologic changes and a score was assigned from 0 to 3. Our study proved aq. glycerine to be a superior rehydrant in comparison to conventional wet fixed smears or coconut oil. Also, in studies where cytoplasmic evaluation is required, coconut oil can be used due to its superior quality.

Comparative Analysis of Cytomorphometric Parameters Among the Coconut Oil, Peanut Oil and Normal Saline in an Air-Dried Smears - A Switch to Routine Wet fixation

Objectives: Rehydration of air-dried exfoliative cytology smears is considered as useful technique to increase the diagnostic accuracy of the air-dried smears and so the study had a point to determine and compare the cytomorphometric parameters of air-dried smears by rehydration using coconut oil, peanut oil and normal saline. Material and Methods: A total of 135 smears were obtained from healthy volunteers and were air-dried. The smears were classified into three groups based on the rehydrating agent: coconut, peanut, and normal saline. The air-dried smears were rehydrated for a period of 5 minutes. And subsequently stained with Papanicolaou stain. The smears were studied under a microscope to evaluate cytoplasm and nuclear details and the quality of the background staining. Scores were assigned to each group and statistically analyzed using the chi-square test. Results: All the studied parameters, such as cytoplasm, nucleus, and background, demonstrated high frequencies of superior scores in the peanut oil group, followed by the coconut oil group and the normal saline group. (p < 0.05) Conclusion: Peanut oil is a rehydrating agent superior to coconut oil and can rehydrate the air-dried smears.

Role of Interphase FISH Assay on Air-Dried Smears in Identifying Specific Structural Chromosomal Abnormalities among Pediatric Patients with Acute Leukemias.

The online version contains supplementary material available at 10.1007/s12288-023-01699-2.

Cytological features of oral malignant lymphoma in scraping liquid-based cytology: Cases of plasmablastic lymphoma and anaplastic lymphoma kinase-positive anaplastic large cell lymphoma

The main purpose of cytological examination in the oral region is to screen for squamous cell carcinoma or intraepithelial neoplasms; thus, the background tends to be considered a deterrent for microscopy. From this perspective, liquid-based cytology (LBC) is favorable for preparing clear samples with few backgrounds. However, background hemocytes are sometimes of critical importance in the diagnosis. We report two cases of oral malignant lymphoma, plasmablastic lymphoma, and anaplastic large cell lymphoma in which careful observation of the background in scraping LBC sample contributed to the early diagnosis. Atypical lymphoid cells were observed only in a very small part of the LBC samples from the presented patients; however, cytological findings, such as large lymphoid cells with outstanding nucleoli, large mitotic cells, or intermediate-to-large lymphoid cells with pleomorphic nuclei were sufficient for obtaining a cytological diagnosis of malignant lymphoma. Although the number and cell size of leukocytes in LBC with Papanicolaou staining were significantly different from those in air-dried conventional smears with Romanovsky staining, which are commonly preferred for the discrimination of hemocytes, the corresponding cytological features could be observed. Therefore, attention should be paid to the background as well as squamous epithelium to prepare for such unexpected cases. The LBC examination with Papanicolaou staining alone can suggest the possibility of malignant lymphoma.

Cytomorphological Comparison between Well-fixed Smear and Air-dried Smear

INTRODUCTION: Cervical cancer is the leading cause of cancer related death, especially in developing countries. The aim of this study was to compare well-fixed smear and air-dried smear. MATERIALS AND METHODS: A prospective study was conducted in Kasturba Medical College, Manipal, India, from paired cervical smears made from 200 women, who attended OPD of Obstetrics and Gynecology during a period of 3 months from November 2010 to January 2011. Routine smears were fixed immediately in 95% of ethanol and stained by Pap stain. Air-dried smears were rehydrated by immersing the slides in normal saline for 30 seconds, fixed in 95% of ethanol and stained by Pap stain. All slides were examined and assessed for various cytological parameters by the pathologist. Bethesda system was followed for reporting the cytology smears. RESULTS: Among the 200 paired smears 199 were negative for epithelial lesion or malignancy. One case (0.5%) had epithelial abnormality in both wet-fixed (WF) and air-dried (AD) smears. However the AD smears showed more number of abnormal cells and cells were of higher grade than the WF smear. Detection rate of Candida was similar (12.5%) in both AD and WF smears. Identification of Trichomonas was also similar (1.5%) in both types of smears. Candida could be more easily identified in AD smears as compared to WF probably because AD had a clearer background. CONCLUSIONS: In air-dried smears, individual cells are clearly seen. The cellular and nuclear size is comparable to tissue sections.

Pros and Cons of Air-Dried and Conventional Wet-Fixed Pap Smears: An Institutional Study

Introduction: Wet-fixed smear study routinely used to identify several pre-cancerous and cancerous tumours. By contrasting cellular preservation and staining quality in rehydration smears with wet-fixed smears, the current study aims to establish the effects of rehydration on air-dried smears. Materials and Methods: One hundred paired samples are compared. Wet fixing was done in the first place. Air-dried samples are rehydrated for 30 s with regular saline and then placed in 95% ethanol for 30 min. Both smears will undergo screening, evaluation and grading for numerous cytomorphological factors after Pap staining. The 2014 Bethesda system was used to report the cervical smear results. Results: When evaluated for numerous cellular properties, statistically significant similarities can be seen between two groups, one with wet-fixed smears and the other with rehydrated air-dried smears. The results of the cytological diagnosis comparison between the two groups can be shown in the agreement. Rehydrated air-dried smear interpretation and staining quality are significantly superior to traditional wet smears. Conclusions: Rehydration air-dried techniques can be used instead of traditional wet fixation methods and are as effective when used alone or with traditional wet-fixed smears.

Prolonged Duration of Air-Dry Fixation of Cervical Smears Produces Superior Cytomorphological Staining Quality over Conventional Wet-Fixed Smears

Introduction: Immediate fixation of smears in 95% alcohol for Pap staining is commonly used for cytopathological diagnosis of cancers and other diseases. Few research studies have investigated the comparative outcomes of alcohol wet fixation and rehydration of air-dried smears, indicating that air-dried rehydrated (ARF) smears are viable alternative to wet-fixed (WF) smears. However, there are no or few investigations on the effects of long durations of air-drying fixation on cytomorphological staining quality. Materials and Methods: 124 cervical smears were obtained from the Family Planning Unit of Komfo Anokye Teaching Hospital in Kumasi, Ghana. Quadruple smears were WF and air-dried for 2 h, 4 h, and 8 h prior to rehydration with normal saline and fixation (ARF). All the smears were stained with Papanicolaou stain, examined microscopically for their cytomorphological features, and scored. Cytomorphological scores were statistically analyzed using SPSS software. Results: No significant difference in cytolysis, cell border, nuclear border, chromatin, and cellularity between WF and ARF was observed. However, significant differences in cytoplasmic staining quality (p value <0.001) and the absence of RBCs (p value <0.001) were observed in the 4-h ARF. The absence of RBCs in the ARF smears rendered a clearer background than in the wet fixation. Conclusions: ARF, Pap-stained smears showed comparably superior cytomorphological features to those of WF smears. The 8-h ARF smears produce crispy chromatin and excellent background, making them suitable for bloody cytological samples.

Fine Needle Aspiration Cytology of Lymphnodes: A Hospital Based study in a Tertiary Health Care Center in Southern Odisha

Introduction: FNAC is widely accepted the most accurate sensitive, specific, cost-effective minimally invasive OPD technique to establish the basic diagnostic information not only to relieve the anxiety of the patients but also to provide materials for microbial culture, and decrease unnecessary surgery from the visibly accessible superficial masses. It also distinguishes a benign from a malignant entity that can obtain tissue for immuno-phenotyping &amp; molecular studies and obtain cellular &amp; genetic material for storage also useful for staging and follow up including response to treatment in patients with known malignancies. Our study was conducted on 796 Outpatients including all age groups &amp; sexes presenting with palpable enlarged lymph nodes not only for early diagnosis &amp; treatment but also to reveal the burden of various categories of diseases in tribal areas where our institution is situated. Material &amp; methods: FNAC was performed under aseptic precaution using a 22 to 24 Gauge needle attached to a 20CC disposable syringe. The aspirated material after smearing was treated with 95% ethyl alcohol for fixation and stained with H&amp; E and Papanicolaou stain &amp; air-dried smears are stained with Leishman stain. Modified Z-N stain was done wherever necessary followed by diagnosis based on clinic cytomorphological features. Result: Out of 796 cases the most common cause of lymphadenopathy in our study was found to be Tubercular lymphadenitis with 360 (45.22%) cases, the second most frequent entity was reactive lymphadenitis in 277(34.79%) cases followed by metastatic lesions in 47 (5.90%) cases and lymphoma 17 (2.13%) cases. Conclusion: FNAC is a simple, safe, reliable, and inexpensive method for early detection of various non-neoplastic (lymphadenitis) &amp; neoplastic lesions (lymphoma &amp; metastatic carcinoma) of enlarged lymph nodes which helped diagnose approximately 99% of cases.

Usefulness and Limitations of Cryopreservation for Immunocytochemical Staining of Canine Cytological Specimens for Detection of Cytokeratin and Vimentin.

Immunocytochemistry is an advanced diagnostic tool for identifying the origin of tumor cells. This study aimed to highlight the usefulness of cryopreserved, air-dried cytological samples in detecting cytokeratin and vimentin. Air-dried cytological smear samples were prepared from a total of 39 resected canine tumors and stored in a medical freezer without fixation. The duration of cryopreservation ranged from 2 to 56 months. The same tumors were processed for routine histopathological examination. Based on the morphological diagnosis, cryopreserved FNA smears from epithelial tumors were stained by enzymatic immunocytochemistry (ICC) for cytokeratin; those from mesenchymal and melanocytic tumors were stained by ICC for vimentin. To ascertain the positivity of tumor cells to the selected markers, tissue paraffin-embedded sections were also stained by immunohistochemistry (IHC) for the same markers. Immunoreactivity for cytokeratin was detected in cryopreserved cytological smears for a maximum of 46 months. Immunoreactivity for vimentin was clearly detected for 33 months. Smears stored at room temperature for 1 week did not show any signals under immunocytochemical examination. Thus, immunocytochemistry for cytokeratin and vimentin can be safely applied to air-dried smears cryopreserved in a freezer for at least 33 months.

Cytopathological profile of patients with hepatic lesion: A hospital-based cross-sectional study from North-east India

Background: Cancer is not a notifiable disease in India. The data pertaining to liver cancer and hepatic lesions are very limited in India. Aims and Objectives: The current study was intended to observe the proportion of different types of liver lesions reported from private hospitals in Mizoram and also to study the association of liver malignancies in relation to gender and age. Materials and Methods: This is a cross-sectional study design. The data were collected from the cytopathology data base of a private laboratory (Genesis Laboratory) in Aizawl, Mizoram. Data regarding 204 patients with hepatic lesions, who have been referred from three private hospitals to a private laboratory in Mizoram from January 15, 2018, to November 31, 2021, were included in the study. Details regarding cytopathology reports of image-guided fine needle aspiration cytology of liver mass, age, and gender were collected. Air-dried smears received at a private laboratory were routinely stained with Leishman-Giemsa and Papanicolaou stains, Ziehl-Neelsen staining was done wherever needed. From the cytopathological diagnosis made, the frequency and patterns of various hepatic lesions were then analyzed using SPSS Version 22. Results: Out of 204 cases, 15 (7.35%) cases were reported as inadequate samples and 4 cases (1.96%) were inconclusive in diagnosis. Hence, remaining 185 study participants were included in the final analysis. The mean age was 58. Majority 122 (65.40%) were males and in the age group 41–60 years (49.19%). Majority of the lesions were malignant 145 (78.37%) and remaining 40 (21.62%) were benign lesions. The most common malignant lesion found was hepatocellular carcinoma (HCC) 87 (47.02%), followed by metastatic carcinoma 31 (16.75%). Liver malignancies were found to be higher among males (74.5%) as compared to females (14.3%), but were not statistically significant (P=0.08). The proportion of males (52.5%) versus females (36.5%) diagnosed with HCC was statistically significant (P=0.03). Conclusion: The most commonly reported liver malignancy was HCC. The proportion of males diagnosed with HCC was higher than females in the present study.

Cytodiagnosis of rhinoscleroma and its morphological clues: Our experience with four cases

Rhinoscleroma, a chronic progressive inflammatory disease of the upper respiratory tract, has a prolonged course and passes through three overlapping clinicopathologic stages. The histopathology findings are well illustrated in the literature; however, there is a paucity of reports explaining cytomorphological features of rhinoscleroma. It is the proliferative stage when disease presents as a nodular mass which can be targeted for cytological study. Here, the authors report fine-needle aspiration (FNA) cytology of four cases of rhinoscleroma who presented with intranasal firm, nodular lesions; its cytology revealed numerous, large, singly scattered histiocytes in a dense inflammatory background of lymphocytes, plasma cells, and few eosinophils. The microorganisms were visualized as intracytoplasmic loosely clustered, short rods within the histiocytes (Mikulicz cells) in May-Grunwald-Giemsa (MGG) smears. FNA sampling of rhinoscleroma in its nodular stage can yield classic cytomorphology with demonstration of organisms on air-dried smears which can further substantiate the cytodiagnosis. Thus, high index of suspicion and diligent search can aid in early and accurate diagnosed even on cytology with the help of conventional wet fixed Pap and air-dried MGG stain; both being complementary to each other and further can be supported with special stains such as periodic acid–Schiff stain or Gram whenever required.

Diff-Quik: A reliable stain for rapid onsite evaluation and differentiation of round cells in semen

Objectives: Identification of immature germ cells and leukocytes and their differentiation pose difficulty in interpretation. In this study, Diff-Quik (DQ) and Papanicolaou (Pap) are used to differentiate between these two, as they are recommended by the WHO, cost-effective and are easy to use. Materials and Methods: This study attempts to differentiate between immature germ cells and leukocytes in semen; compare the usefulness of two stains –Pap and DQ; evaluate the possible associations between the total sperm count and the round cell count; the total sperm count and the presence of round cells and inflammatory cells; the occupation and the sperm concentration; and the occupation and the round cell count. Results: A total of 60 semen samples were included in the study. Two smears were made for each patient; DQ stain was used to stain the air-dried smears and Pap stain for alcohol-fixed smears. The whole procedure of DQ staining takes only 30 s. The smears were compared. There was good concordance between DQ and Pap stains in the detection of inflammatory cells and immature germ cells (P = 0.000). There was no association between total sperm count and round cell count (r = −0.086 and P = 0.520); and between the occupation and the sperm concentration or the round cell count. Conclusion: There was an inverse relationship between total sperm count and presence of immature germ cells (r = −0.291). There was no association between round cell count and presence of immature germ cells (r = 0.148 and P = 0.269), total sperm count and presence of inflammatory cells, round cell count, and presence of inflammatory cells. DQ stain is easy to use, convenient, and rapid. There is good concordance between DQ and Pap stains in the detection of inflammatory cells and immature germ cells. Total sperm count is independent of the round cell count or the presence of inflammatory cells/immature germ cells. In most of the cases, the round cells were the inflammatory cells. Morphological interpretation can be done even in non-liquefied samples.

Fine Needle Aspiration Cytology of Lymphnodes: A Hospital Based study in a Tertiary Health Care Center in Southern Odisha

Introduction: FNAC is widely accepted the most accurate sensitive, specific, cost-effective minimally invasive OPD technique to establish the basic diagnostic information not only to relieve the anxiety of the patients but also to provide materials for microbial culture, and decrease unnecessary surgery from the visibly accessible superficial masses. It also distinguishes a benign from a malignant entity that can obtain tissue for immuno-phenotyping &amp; molecular studies and obtain cellular &amp; genetic material for storage also useful for staging and follow up including response to treatment in patients with known malignancies. Material &amp; methods: FNAC was performed under aseptic precaution using a 22 to 24Gauge needle attached to a 20CC disposable syringe. The aspirated material after smearing was treated with 95% ethyl alcohol for fixation &amp; stained with H&amp; E and Papanicolaou stain &amp; air-dried smears are stained with Leishman stain. Modified Z-N stain was done wherever necessary followed by diagnosis based on clinic cytomorphological features. Result: Out of 796 cases the most common cause of lymphadenopathy in our study was found to be Tubercular lymphadenitis with 360 (45.22%) cases, the second most frequent entity was reactive lymphadenitis in 277(34.79%) cases followed by metastatic lesions in 47 (5.90%) cases and lymphoma 17 (2.13%) cases. Conclusion: FNAC is a simple, safe, reliable, and inexpensive method for early detection of various non-neoplastic (lymphadenitis) &amp; neoplastic lesions (lymphoma &amp; metastatic carcinoma) of enlarged lymph nodes which helped diagnose approximately 99% of cases.

Cytomorphology of lymphadenopathy on fine needle aspiration cytology

Introduction: FNAC is widely accepted the most accurate sensitive, specific, cost-effective minimally invasive OPD technique to establish the basic diagnostic information not only to relieve the anxiety of the patients but also to provide materials for microbial culture, and decrease unnecessary surgery from the visibly accessible superficial masses. It also distinguishes a benign from a malignant entity that can obtain tissue for immuno-phenotyping &amp; molecular studies and obtain cellular &amp; genetic material for storage also useful for staging and follow up including response to treatment in patients with known malignancies. Our study was conducted on 796 Outpatients including all age groups &amp; sexes presenting with palpable enlarged lymph nodes not only for early diagnosis &amp; treatment but also to reveal the burden of various categories of diseases in tribal areas where our institution is situated. Material &amp; methods: FNAC was performed under aseptic precaution using a 22 to 24Gauge needle attached to a 20 CC syringe (disposable). The aspirated material after smearing was treated with 95% ethyl alcohol for fixation &amp; stained with H&amp; E and Papanicolaou stain &amp; air-dried smears are stained with Leishman stain.

Usefulness of cellblock preparation in fine needle aspiration for the diagnosis of thyroid nodules.

Thyroid nodules are common in the general population. The current diagnostic method for nodules is the ultrasound guided fine needle aspiration (US-FNA). The aim of the study was to evaluate the usefulness of cellblock preparation in addition to routine US-FNA in the diagnosis of thyroid nodules. A retrospective study of patients with thyroid nodules submitted to US-FNA, with collection of material using both smears and cellblock preparation. Two air-dried smears were prepared for each nodule. After centrifugation, the residual aspirate in the syringe and needle was processed as a standard histology specimen (cellblock). Then a pathologist reviewed the smears and cellblock slides of each case. A total of 12.360 thyroid nodules were submitted to US-FNA. Cellblock preparation was performed in 153 (1.2%) in addition to smears. Among the satisfactory cellblocks (80.5%, 120), 31.7% (38) provided additional morphological information in comparison with smears alone. No significant differences were found between the smear and the combined smear and cellblock evaluation concerning the number of unsatisfactory (12.1% vs. 11.4%, p=.85) and indeterminate (27.5% vs. 24.2%, p=.52) results. Overall, 10 samples (6.7%) had their diagnosis changed after cellblock evaluation, nine of them due to immunohistochemical studies. Immunohistochemistry confirmed parathyroid origin of the nodule in six cases. Cellblocks did not contribute to increase cellularity of the samples or to reduce indetermined results of FNA of thyroid nodules. Immunohistochemistry was essential to characterize rare cases without follicular histogenesis. Cellblock must only be prepared when considering performing immunohistochemistry.

Feasibility of using digital confocal microscopy for cytopathological examination in clinical practice

Optical imaging modalities are emerging as digital microscopy tools for tissue examination. The investigation of these techniques for potential applications in anatomic pathology practice has focused primarily on surgical pathology and has not included cytopathological specimens. We evaluated the feasibility of using digital confocal microscopy (CM) to examine cytopathological specimens. Smears and cell suspensions collected in RPMI solution were prepared from tissue scrapes obtained from surgical resections of breast, lung, liver, and kidney. Air-dried smears and cell pellets obtained from centrifugation of the cell suspensions were stained with 0.6 mM acridine orange and imaged with a CM platform. After completion of imaging, the smears were stained with Diff-Quik (DQ), and cell pellets were routinely processed, embedded in paraffin wax, cut, and stained with hematoxylin and eosin (H&E). We evaluated the mean time to acquire digital CM images; quality of images based on the extent of tissue recognition (0%, grade 0; 1–19%, grade 1; 20–50%, grade 2; >50%, grade 3); comparison of images with DQ- and H&E-stained specimens; and ability to make specific diagnoses. We imaged 91 smears and 52 cell pellets and acquired digital CM images within 2–3 min, with 92% and 88% of images, respectively, from smears and cell pellets showing grade 3 quality. On the basis of CM images, 8 smears (9%) and 7 cell pellets (14%) were categorized as benign, and 83 (91%) and 45 (88%), respectively, as malignant. Specific diagnoses were made by using digital CM images of smears and cell pellets that matched accurately with corresponding DQ- and H&E-stained preparations. The results of our first feasibility study clearly indicated the utility of CM as a next-generation digital microscopy tool for evaluating cytology specimens. Prospective clinical studies are warranted for validating our findings for potential incorporation into cytopathological clinical practice.

Evaluation of immunocytochemistry on destained giemsa stained smears as an alternative to conventional technique.

Protocol for immunocytochemical (ICC) staining in May-Grünwald Giemsa (MGG)-stained smears has been difficult to establish. It is the need of the hour to be able to use prestained slides for ICC in specific cases to deliver timely diagnoses and reduce inconvenience to patients. To evaluate and compare the use of MGG-stained smears for the purpose of ICC, after de-staining and saline rehydration to that of routine standard ICC. A prospective study was conducted on 40 FNAC samples: 25 cases of breast disease and 15 cases of reactive lymphoid hyperplasia known to express pancytokeratin and leukocyte common antigen (LCA)/CD45, respectively. Air-dried smears of each case were stained by standard MGG stain and after the report was dispatched, one smear was selected and sent for ICC. The smears were analyzed to determine the overall result and grade each smear semi-quantitatively with respect to staining-intensity, stain-localization, staining-uniformity, counter-staining, and background-staining. The proposed protocol was inferior to conventional ICC in all the parameters, more pronounced in pancytokeratin than LCA/CD45. Only 8% of air-dried smears stained for pancytokeratin showed optimal stain intensity (as opposed to 44% of wet-fixed smears), whereas only 14.3% of air-dried smears were optimally stained for LCA (as opposed to 85.7% of wet-fixed smears). The proposed protocol of de-stained Giemsa smears as an alternative to conventional technique for ICC was unsuccessful in giving satisfactory results.

Immunohistochemistry Applied to Breast Cytological Material.

Fine-needle aspiration biopsies (FNABs) of the breast are minimally invasive procedures enabling the diagnosis of suspicious breast lesions. Unfortunately, they are often perceived as inferior to core-needle biopsies, namely because they are supposedly unable to differentiate between high-grade ductal carcinoma in situ and invasive carcinoma or provide material for ancillary testing. Several studies have shown, however, that FNAB samples, when handled properly, are indeed capable of providing sufficient and adequate material for ancillary testing, namely immunocytochemistry (ICC). We reviewed the published literature regarding the use of ICC for both diagnostic and theranostic uses in the different types of cytological samples obtained from FNABs of the breast, including smears, liquid-based cytology samples, and cellblocks. We found that p63 and 34βE12 show promise in aiding in the differential diagnosis between in situ and invasive lesions and that most other diagnostic markers may be used as in tissue. Regarding theranostic ICC markers, results vary between publications, but with care, these can successfully be performed in cytological samples. Air-dried smears should be avoided, and cellblocks are overall more versatile than cytology slides, enabling the evaluation of not only hormonal receptors and HER2 by ICC, but also of Ki-67. Particular attention should be paid to fixation and antigen retrieval procedures in all cases. We recommend that laboratories without experience perform short validation runs before adopting these techniques into clinical practice.