Routine Tissue Processing Research Articles

Tissue Processing using Microwave Oven: A Boon for Histology Slide Preparation

Background: Tissue processing is an important step in histology laboratories. Routine tissue processing is time-consuming. Due to the same time factor, microwave tissue processing is now increasingly being used in many histology and pathology laboratories. Objective: To compare the time taken to process the tissues using the routine conventional and microwave methods. Methods: The study was conducted in the Histology laboratory. 100 slides (50 conventional method + 50 microwave method) were processed using both methods, sectioned, stained, and analyzed by an independent observer who was unaware of the processing method. Results: The study showed a significant decrease in processing time by the microwave method (p-value < .0001). Conclusion: The present study concluded that the time taken for dehydrating, clearing, and embedding the tissues in paraffin wax was found to be considerably lower than that taken for the conventional method of tissue processing. Chloroform was used here as a clearing agent, and it had desirable effects. KEYWORDS: Microwave tissue processing, Conventional tissue processing.

Determination of local expressions of IGF-1, LC3B and NF-kB in white muscle disease in lambs by immunohistochemical method

White muscle disease (WMD) is also known as Stiff Lamb Disease or Nutritional Muscular Dystrophy. Selenium and/or Vitamin E deficiency constitutes the etiology of the disease. This study aimed to immunohistochemically evaluate local protein expressions of Nuclear factor kappa B (NF-kB), Insulin-like growth factor-1 (IGF-1) and Microtubule-related protein 1A/1B-light chain 3 beta (LC3B) in WMD. The material of the study consisted of 15 WMD, and 6 healthy lamb heart samples. The heart tissues of the autopsied lambs were subjected to routine tissue processing and paraffin blocks were obtained. Then, it was stained with Hematoxylin-Eosin and immunohistochemical methods. Control group lambs had normal macroscopic appearance. Macroscopically, hyaline degeneration and zenker’s necrosis, calcification areas were observed in WMD tissues. Microscopically, degenerative and necrotic muscle fibers, calcification areas, fibrosis, mononuclear cell infiltrates and macrophage infiltrates were detected in WMD heart tissues. Immunohistochemically, significant increases were detected in IGF-1 (p

Histopathological changes in the placentas of pregnant women with Gestational Diabetes Mellitus

Objectives: Placental damage might be the primary underlying cause of the high rates of fetal adverse outcomes in cases with gestational diabetes mellitus (GDM). Histopathological changes were considered to be associated with GDM in the literature. However, there are also studies showing no changes in the placenta in some gestational diabetic women. The pathophysiology of these placental alterations remains elusive. Therefore, this study aimed to examine the histopathological changes in the placentas of patients with GDM and compare them with the placentas of the healthy control group.
 Material and methods: This experimental non-randomized case-control study was conducted with 40 GDM patients and 40 healthy pregnant women at Dicle University Faculty of Medicine between March 2023 and December 2023. Placental tissues obtained following cesarean delivery were subjected to routine tissue processing. Placental samples were stained with hematoxylin-eosin and examined histopathologically using a light microscope.
 Results: When histopathological findings were compared between the GDM and control groups, villous degeneration, congestion in blood vessels, fibrinoid accumulation, inflammation and intervillous hemorrhage were higher in the GDM group than in the control group (p<0.001).
 Conclusion: The intricate relationship between GDM and placental pathology underscores the profound impact of metabolic changes on placental structure and function throughout pregnancy. Understanding these pathological changes is crucial for elucidating the progression of GDM and its implications for maternal and fetal health, highlighting the importance of comprehensive placental analysis in managing pregnancies complicated by GDM.
 Keywords: Gestational diabetes, hematoxylin-eosin staining, histopathological examination

Standardization of honey as a tissue fixative for histopathology: A morphometric study

Background: Tissue fixation is a crucial step to preserve the tissues in a life-like state with minimal disruption to its cellular and chemical composition for histopathological examination. The search for an effective alternate tissue fixative to the routinely used formaldehyde has gained interest as constant exposure to formaldehyde has proven to be toxic. Honey, an organic substance with high acidity and hygroscopic nature, exhibits tissue fixative properties and has been used in the present study. The present study aimed to standardize honey as a tissue fixative for histopathology by comparing it with formalin. Materials and Methods: In vitro study Oral tissue samples of goat were fixed in 10% honey and 10% formalin solution, respectively, for 24-48 h, followed by routine tissue processing and microscopic examination of 37 slides per group. 2200 epithelial cells (1100 per group) were selected for the computer-aided morphometric image analysis (Fiji-Image J) by three observers. Cell area (CA), cell perimeter (CP), nuclear area (NA), nuclear perimeter (NP), cytoplasmic area (Cyt A), and nuclear-cytoplasmic ratio were the parameters studied. Mann-Whitney U-test (STATA/IC version 16) for inter-group comparison was done and P < 0.05 was considered statistically significant. Results: The probability of epithelial cells in the honey-fixed group to have greater NA, NP, and N/C ratio was about 50%-60%. The probability of epithelial cells in formalin-fixed tissues to have greater CA, CP, and Cyt A was about 70%. Conclusion: Honey is a better nuclear fixative than formalin. Cytoplasmic shrinkage of epithelial cells should be taken into consideration while fixing tissues with honey.

Assessment of the Effects of Two Different Doses of Methotrexate in Rat Ovarian Tissue: A Histological Study

Objective: The use of methotrexate (MTH), a drug commonly prescribed to treat rheumatoid arthritis, cancer, and certain autoimmune diseases, has been linked to early ovarian failure and infertility in women when used in varying amounts and durations. In this study, the possible effects of MTH exposure at different doses on rat ovary were investigated histologically and immunohistochemically. Materials and Methods: A total of 21 female rats, Wistar albino, were utilized in the investigation. The control group (n = 7) was not subjected to any treatment. The 10 mg/kg MTH group (n = 7) and the 20 mg/kg MTH group (n = 7) were given intraperitoneal injections of 10 mg/kg and 20 mg/kg methotrexate, respectively. Five days after injection, the ovarian tissues were taken out of the anesthetized rats and subjected to routine histological tissue processing. The histological evaluation was performed with hematoxylin and eosin (H & E) and Masson's trichrome (MT) on paraffin block sections. Additionally, immunohistochemical staining was performed to determine AMH immunoreactivity. Results: In both MTH groups, histopathological changes were observed, particularly in the 20 mg/kg group, and these included the presence of areas of edema as well as dilated or congested blood vessels. Furthermore, the 20 mg/kg MTH group exhibited increased fibrotic tissue in the ovarian medulla. The number of follicles and AMH immunoreactivity were observed to be reduced in the MTH groups, with the reduction reaching a statistically significant level in the 20 mg/kg MTH group ovary. In terms of AMH immunoreactivity, it is demonstrated that there is no statistical significance between the control and 10 mg/kg MTH groups. Conclusion: The dose of MTH should be considered carefully, as increasing the dose may lead to deterioration of ovarian histopathology and a reduction in the reserve of the ovary.

Investigation of the Sox-9 and Caspase-6 Immune Activity in Placentas of Pregnant Women with GDM

Aim: In this study, we investigated the immunohistochemical staining of Sox-9 and Caspase-6 expression in placentas of pregnant woman with gestational diabetes mellitus (GDM). Material and Methods: Placentas of 20 healthy and 20 women with GDM were processed for routine histological tissue processing. The biochemical and clinical parameters of patients were recorded. Placentas were stained with hematoxylin-eosin and Sox-9 and Caspase-6 immunostaining. Results: In control group, Sox-9 expression was negative in decidual and connective cells and endothelial cells. In GDM group, Sox-9 expression was increased especially in the decidual cells. For the Caspase-6 expression, Caspase-6 reaction was mainly in maternal region in control group. In GDM group, Caspase-6 reaction was increased in decidual cells, in endothelial cells and in the syncytial nodes. Conclusion: Expression of Sox-9 transformed the decidua cells and lead to apoptotic pathway via Caspase-6 expression.

Some histological and biochemical evaluation on commercial hair dyes

The use of hair dyes as a means of beautification is currently increasing. This study aimed at evaluating the toxicity of different commercial hair dyes on the histology of the skin and biochemical parameters of the liver of Albino rats. This study was carried out in the Pathology Department, University of Calabar Teaching Hospital Calabar and College of Medical Sciences, University of Calabar, Calabar between May, 2019 and October, 2019. Four different colours of hair dyes were obtained from a cosmetic store in Watt daily market in Calabar, Cross River State. Twenty male Albino rats with average weight of 55g were used for this study. The rats were randomly divided into five (5) groups of four (4) rats each. Group 1 to 4 served as the test groups while group 5 served as the control group. Different colours of hair dye were applied topically on selected area of the back of Albino rats daily for 30 days after which, the rats were sacrificed using chloroform inhalation procedure. The skin of the rat was harvested for histology while heart puncture was used to collect 5 milliliters of blood from each rat for biochemical analysis. Plasma Alkaline phosphatase (ALP) was determined using Kind and King's method. Plasma Alanine amino transferase (ALT) and Aspartate Amino transferase (AST) were determined by Reitman and Frankel method. Serum protein (TP) was determined by Biurette's method while bilirubin was estimated using Powell's method. Liver tissues were processed using routine paraffin wax tissue processing method for histological analysis. The serum levels of ALP, AST, TP, Total bilirubin (TB) and conjugated bilirubin (CB) in all the test groups were significantly increased when compared with the control (P=0.001). Histological evaluations indicated evidence of cellular injury in all the test groups. Prolonged use of commercial hair dyes caused adverse effects to the skin and liver as seen in the Albino rat used in this study.

Investigation of the Sox-9 and Cited-1 Immune Activity in Placentas of Women with Placenta Accreta

Aim: In this study, we investigated the immune activity of the Sox-9 and Cited-1 in women with placenta accreta. Material and Methods: 20 healthy and 20 placenta accreta were processed for routine histological tissue processing. Placentals samples were dissected and fixed in 10% formaldehyde solution. Samples were embedded in paraffin blocks. Clinical and biochemical parameters were recorded. Placental sections were cut from paraffin blocks and stained with Sox-9 and Cited-1 immunostaining. Results: In our study, control group showed negative Cited-1 expression in decidual cells, root villi and connective tissue areas in general. Placenta accreta group showed increased Cited-1 expression in degenerated decidual cells, fibroblastic cells and endothelium. In control group, Sox-9 expression was negative in the syncytial knots, in the vascular endothelial cells. In placenta accreta group, Sox-9 reaction was positive in the root villi, in the blood vessels, in the connective tissue. Conclusion: It was observed that the Sox-9 reaction was increased and inflammation was induced, depending on the differences in decidual cells, in the syncytial area and in the vascular endothelium in in placentas of women with placenta accreta. It is thought that Sox-9 signaling processes are being determined and Cited-1 may be stimulants that affect cell proliferation and angiogenesis regulation and affect placental development.

Role of cited-1 and caspase-6 expression in HELLP syndrome.

In this study, we investigated the immunohistochemical staining of cited-1 and caspase-6 expression in the placentas of pregnant women with HELLP syndrome. Placentas of 20 normotensive patients and 20 women with HELLP syndrome were processed for routine histological tissue processing. The biochemical and clinical parameters of patients were recorded. Placentas were stained with hematoxylin-eosin and cited-1 and caspase-6 immunostaining. Placentas of normotensive patients showed normal histology. Placentas of women with HELLP syndrome showed degenerated cells, hyalinization and vacuolization. Cited-1 expression was negative in normotensive group; however, it was increased in HELLP group, especially in decidual cells, endothelial cells and other placental cells. Caspase-6 expression was negative in placental structures of normotensive groups. However, it was intense in decidual cells, vacuolar and hyalinized areas, inflammatory cells and connective tissue cells in HELLP group. Cited-1 and caspase-6 are a marker in determining the severity of HELLP syndrome.

An investigation of the potential effects of amitriptyline on polycystic ovary syndrome induced by estradiol valerate.

Polycystic ovarian syndrome (PCOS) is frequently observed in adolescent women and usually progresses with depression. The aim of this study was to examine the effects of amitriptyline (Ami), a drug used in the treatment of depression, in individuals with PCOS. Forty 12-week-old female Wistar albino rats were randomly divided into five groups: control, sham, PCOS, Ami, and PCOS + Ami. To induce the syndrome in the PCOS groups, a single dose of 4mg/kg estradiol valerate was administered by intraperitoneal injection; 10mg/kg Ami was administered by intraperitoneal injection for 30days in the Ami groups. After 30days, all the animals were sacrificed and blood, ovary, and brain tissues were collected and subjected to routine tissue processing. Stereological, histopathological analyses were performed on the ovarian sections, while luteinizing hormone (LH), follicle-stimulating hormone (FSH), catalase (CAT), and superoxide dismutase (SOD) levels were investigated in blood samples. The volume of the corpus luteum and preantral follicles increased in the PCOS group, while a decrease was determined in the number of antral follicles using stereological methods. Biochemical analysis revealed that FSH levels increased and CAT enzyme levels decreased in the PCOS group. Significant morphological changes were observed in ovaries from the PCOS group. The volume of the corpus luteum in the PCOS + Ami group decreased compared to the PCOS group. Serum FSH levels decreased in the PCOS + Ami group, while CAT enzyme levels increased compared to the PCOS group. Degenerative areas were also seen in the PCOS + Ami group ovaries. Ami administration was unable to sufficiently ameliorate the morphological and biochemical changes caused in the ovarian tissues by PCOS. In addition, this study is one of the few studies examining the effects of amitriptyline, an antidepressant frequently used in depression treatment of individuals with PCOS. We also observed firstly that use of amitriptyline caused PCOS-like ovarian morphology in healthy rat ovaries, while it had a healing effect by volume decreasing of cystic structures in the ovary with PCOS.

Role of SOX9 and Hif-1α expression in placentas of patients with HELLP.

In this study, we investigated the immunohistochemical staining of SRY-box transcription factor 9 (SOX9) and Hif-1α expression in placentas of pregnant woman with hemolysis, elevated liver enzymes and low platelets (HELLP) syndrome. Placentas of 20 normotensive and 20 women with HELLP syndrome were processed for routine histological tissue processing. The biochemical and clinical parameters of patients were recorded. Placentas were stained with hematoxylin-eosin and SOX9 and Hif-1α immunostaining. Normotensive placentas showed normal histology of placenta, however placentas of HELLP syndrome showed intense thrombosis, thinning of the villi membrane and vascular dilatation. In placentas of normotensive patients, SOX9 reaction was immunohistochemically negative, however placentas of HELLP group showed SOX9 expression in decidual cells, and syncytial regions of floating villi and inflammatory cells. In placentas of normotensive patients, Hif-1α reaction was mainly negative in vessels and connective tissue cells. Placentas of HELLP group showed increased Hif-1α expression in decidual cell and especially inflammatory cells in the maternal region. Hif-1α and SOX9 proteins can be used as a marker to show severity of preeclampsia and regulation of cell proliferation and angiogenesis during placental development.

Qualitative assessment of Aloe vera as natural tissue fixative: An institutional study.

The art of microscopy is only appreciable in well-fixed tissues. We conducted this study to determine the effectiveness of Aloe vera as a tissue fixative and compare its results with the natural fixatives already studied in the literature. A pilot study was tried out using commercially available fresh chicken and fish with Aloe vera, and then after getting the promising results similar study protocol was carried out using 10-autopsied human tissue. Four natural fixatives-30% jaggery solution, 20% honey solution, 20% sugar solution, 20% Aloe vera solution and 10% formalin were used for fixation in the study. Fixation of tissues was carried out at room temperature for 24 h. All pre- and postfixation measurements were recorded using stereomicroscope and its software. The difference between pre-and postfixation was calculated and later, all pieces were kept for routine tissue processing followed by routine staining. The tissue sections were assessed for quality, and the whole procedure was blinded among three oral pathologists who scored them. The mean percentage of shrinkage in each bit with different reagents was calculated. The shrinkage seen with 10% formalin and 20% Aloe vera were more likely similar. Among all the natural fixatives, qualitatively also Aloe vera excelled and its results were comparable to that of formalin. The use of Aloe vera in the present study as fixative is the first of its kind, as exhaustive search of the literature shows only its use as transport media in dentistry.

Effect of miRs-17/20 on vasospasm in subarachnoid hemorrhage model of rats

Aim: To investigate the effects of melatonin and miRNA-17/20 administration on vasospasm and vascular damage on the bacillary artery in the Subarachnoid hemorrhage (SAH) model of rats.Methods: Rats were divided into 6 groups: Sham, SAH, SAH+NegmiRNA, SAH+MEL, SAH-miRs-17/20 group, SAH+MEL+miRs-17/20. For creating the SAH model the skin was cut with a vertical incision in the anterior region of the head.120 µL of fresh non-heparinized autologous arterial blood collected from the tail artery was injected into the prechiasmatic cistern under aseptic conditions. All steps in the Sham were the same as in the SAH group, except for blood injection. In the SAH+NegmiRs-17/20, miRs-17/20 miRNA Mimic-Negative Control#1 was administered 1 hour after SAH operation. In the SAH+MEL,10 mg/kg melatonin was administered intraperitoneally 1 hour after the SAH operation. In the SAH-miRs-17/20, mimic-miR-17 and mimic-miR-20 were given intranasally 1 hour after the SAH operation. In the SAH+MEL+miRs-17/20,intranasal mimic-miR-17 and intraperitoneal melatonin were administered 1 hour after the SAH operation. Brain samples, including the bacillary artery, were taken and subjected to routine tissue processing procedures. Vessel samples were evaluated and graded in histological sections stained with the H-E method in terms of vasospasm, edema in the tunica media, and folding of the lamina elastica interna.Results: The co-administration of melatonin and miRs-17/20 reduced the vasospasm and edema formation in the vessel wall. It has also been demonstrated that the application of miRs-17/20 after SAH alone reduces the development of edema in the vessel wall and folding of the internasal lamina elastica due to vasospasm.Conclusion: It has been shown that miRs-17/20 can reduce vasospasm in the vessel wall and prevent vessel damage by reducing edema.

The effects of alpha-lipoic acid (ALA) on the urinary bladder injury in rats exposed to chronic stress: A histochemical study

Objective: In the present study, we aimed to investigate the morphological and biochemical effects of alpha-lipoic acid (ALA) on bladder injury caused by water avoidance stress (WAS) and to show its effect on the number of degranulated mast cells, which increase after stress. Materials and Methods: Wistar albino rats were subjected to WAS and the animals in the treatment group were injected ALA. After the urinary bladder tissues were subjected to routine tissue processing, hematoxylin-eosin staining and periodic acid-Schiff reaction were applied to observe general morphology and acidic toluidine blue method to investigate mast cells. Biochemical assessments of malondialdehyde (MDA) and glutathione (GSH) were also obtained. Transmission electron microscope was used for the ultrastructural, and scanning electron microscope for the topographical analyses. Results: The experiments showed that chronic stress caused injury in the bladder, increased degranulated and total number of mast cells and decreased GSH and increased MDA levels. ALA treatment after WAS ameliorated bladder injury in most areas, decreased degranulated and total mast cell number and increased GSH and decreased MDA levels. Conclusion: It was concluded that ALA can be a useful agent in the treatment of interstitial cystitis.

Histological and histochemical study of large intestine in Camel (Camelus Dromedarius)

Current study was performed to describe the Histohistological and histochemical structures of the large intestine in Iraqi one humped camel (Camelus Dromedarius). Five healthy male camels Approximately 3 to 4 years old were chosen and their specimens from cecum, colon and rectum. The specimens were directly transferred after the animal’s slaughtering to the histology lab at College of Veterinary Medicine for subsequent histological techniques. Tissue specimensfrom various parts of the large intestine (cecum, colon and rectum) were collected and fixed with 10% neutral buffered formalin then subjected to routine tissue processing. Sections of 6 micrometers thickness were prepared and stained with hematoxylin and eosin (H&E) for general histology and Periodic Acid Schiff combine with Alcian blue (AB–PAS), Masson's trichrome as well as Verhoeff stains for histochemical description. Microscopic findings revealed four tunicae constructing the wall of cecum, colon and rectum mucosa, that were from the inner most mucosa, submucosa, muscularis and the outer most either serosa or adventitia. The tunica mucosa characterized by simple columnar epithelium rested on basement membrane and the presence of muscularis mucosa which separated mucosa from the underneath submucosa. Crypts of Lieberkühn and goblet cells were abundant in the colonic epithelium. The submucosa was made up of irregular connective tissue rich with adipose tissue and was supplied well with many blood vessels. Tunica muscularis in all over the three parts of large intestine was made up of inner circular smooth muscle fibers and outer longitudinal smooth muscle fibers. The tunica serosa, and in parts replaced by adventitia, was made up of loose connective tissues.

Is nerve bundle hypertrophy a reliable criteria for diagnosing Hirschsprung disease? A case control study using Calretinin as an adjunct tool for confirming Hirschsprung disease.

Objective: To determine the frequency of nerve bundle hypertrophy in aganglionic segments in relation to the site of biopsy, along with the assessment of value of Calretinin immunostaining in the diagnosis of suspected cases of Hirschsprung disease. Study Design: Cross Sectional study. Setting: Department of Pathology, Pakistan Institute of Medical Sciences (PIMS) Islamabad. Period: September 2018 to March 2019. Material & Methods: After routine tissue processing colonic biopsies were examined for presence of ganglion cells and hypertrophic nerve presence or absence. Diagnosis of Hirschsprung disease was based on the absence of ganglion cells in submucosal and myenteric plexus, the presence or absence of hypertrophic nerves (more than 4 nerves >30 µm thick/×200 field or more than 2 nerves >40 µm thick/×200 field) was also noted in all cases of Hirschsprung disease (aganglionic segments). Calretinin immunostaining was applied to all the cases and controls and findings were recorded as positive or negative staining. Statistical Analysis: Data was analyzed using SPSS version 23. Qualitative data was calculated as frequencies and percentages. Pearson Chi square test was used to establish the association of nerve bundle hypertrophy with the site of biopsy. Results: Total biopsies were 60; 30 each from ganglionic and aganglionic segments. Calretinin sensitivity in our study was 90%, specificity 83.3%. In 30 cases of aganglionosis hypertrophic nerves were present in 13(21.7%) and they were absent in 17 (28.3%). No significant association (p value= 0.447) was seen in nerve bundle hypertrophy and site of biopsy. Conclusion: Calretinin immunohistochemistry can be used as a reliable ancillary technique in the diagnosis of HD. Aganglionosis may not always be associated with submucosal nerve hypertrophy which alone should not be used as a criteria for HD diagnosis but instead adjunct methods like Calretinin immunostaining must be utilized to confirm presence or absence of ganglion cells. There is no association of nerve hypertrophy with site of biopsy.

Rapid Microwave Tissue Processing and Staining Method Using a Kitchen Microwave Oven in Histopathology Laboratory

Background: Histopathological diagnosis of a specimen requires good histoprocessing. It ultimately depends on sample preparation and staining, which is time consuming by routine tissue processing technique. The objective of the present study was to compare the reliability of kitchen microwave for tissue processing and staining with the standard conventional processing and staining in different tissue samples. Materials and Methods: The study sample included 204 specimens from different parts of the body. Three set of tissues were processed and three set of slides from each sample were stained by routine as well as microwave method and the quality of microscopic features was compared. Results: No significant difference was found in nuclear and cellular characters in case of staining by both methods. While, in case of processing, mean scoring for nuclear characters for routinely processed and microwave-stained (RM) slides was 6.95 ± 0.09 (confidence interval [CI] 6.83–0.09), for microwave-processed and microwave-stained (MM) slides, it was 6.82 ± 0.07 (CI 6.74–6.90) by observer 1 (P = 0.068). The mean score of cellular character of RM slides was 7.85 ± 0.06 (CI 7.76) and for MM slides, it was 7.65 ± 0.17 (CI 7.47–7.88) by observer 2 (P = 0.076). The microwave method was better for both processing (P = 0.001) and staining (P = 0.003) in case of small biopsies (P = 0.057). The microwave method was as good as routine method for processing of aorta and staining of breast and kidney tissues (P = 0.23, 0.32, and 0.38, respectively). The time taken by the microwave method is considerably less for both processing and staining. Conclusion: The microwave method requires less time when compared with the routine method, thus reducing the overall turnaround time which ultimately helps in rapid same-day diagnosis and patient management. Microwave can be preferred method for smaller biopsies when compared with large biopsies. Lack of automation and requirement of more manpower remain the major disadvantages for use of microwave method in large turnover histopathology laboratories.

Histological Structure of the Plastinated Kidney Following Deplastination

Plastination is a laboratory process to obtain permanent dry tissue and organ sample. That can be used in the future for investigation and educational purposes. This methodology is based on dehydration and penetration of synthetic substances such as silicon into tissue. In this study, it was aimed to deplastinate previously plastinated kidneys in order to examine them under the light microscope. In this study, 14 sheep kidneys were used, seven samples of control and seven samples of plastination-deplastination (p / d) group. Kidneys in both control and p/d groups were fixed in 10% formalin. The samples in the control group were embedded in paraffin following routine tissue processing protocol. However, the samples in the p/d group were deplastinated in alcohol and methylbenzene and embedded into paraffin. 5 μm thick sections obtained from paraffin blocks were stained with hematoxylin and eosin (H&E), periodic acid-shiff (PAS) and then examined under the light microscope. Typical histological structures were observed in the control group. Small fragments were obtained as it was challenging to obtain sections from the P/d group blocks. Morphological structures were visible with some pseudo degenerations and wrong staining. This study is the first study that demonstrates alcohol and methylbenzene deplastination can be partially successful for evaluating plastinated kidney samples under a light microscope. However, we believe that the kidney may have limitations due to its wide parenchyma compared with literature conclusions. Nevertheless, more studies are required to develop the optimum protocols.

Comparison of homogenization with the GentleMacs Dissociator vs conventional methods for routine tissue processing.

Tissue specimens are valuable materials for microbiological diagnosis. The method of tissue processing can have a significant effect on sensitivity. This study aimed to compare different biopsy processing methods in terms of efficacy and standardization. Pork tissue artificially inoculated with Staphylococcus aureus and Escherichia coli, and samples of infected human tissue were processed by different methods before culture, and the results compared. Bacterial recovery from artificially inoculated pork tissue was significantly higher by homogenization with GentleMacs Dissociator than with sonication. No significant difference was observed between the GentleMacs Dissociator and manual treatment with a scalpel and vortexing. The microbial yield from homogenized human tissues was significantly higher after homogenization with GentleMacs Dissociator than with the conventional method. Homogenization with the GentleMacs Dissociator retrieves bacteria from tissue effectively. Tissue homogenization with the Dissociator is easy and fast to perform and allows for a high degree of standardization.

Spectrum of Morphologic Changes in Histopathological Specimen of Gallbladder in a Tertiary Care Hospital

Background: Diseases of gallbladder are incidentally found and when symptomatic, present with sign and symptoms of cholecystitis and cholelithiasis. Routine examination of gallbladder both grossly and microscopically shows many interesting findings. Aim: The aim of the study was to observe and analyze the diverse morphological and histopathological features of gallbladder specimens following cholecystectomy procedure. Methods: This cross-sectional study was carried out at the Department of Pathology, Gazi Medical College, Khulna; from April 2019 to March 2021 and the sample size was 62. After routine tissue processing and H/E staining all the slides were thoroughly examined under light microscope stains to confirm the diagnosis as well as to evaluate histopathologic characteristics. The data was tabulated and statistical analysis was performed. Results: Out of 62 cases 30.64% samples belonged to age 26-35 years and the mean age was 44.32 years. 69% patients were female with a female to male ratio 2.26:1. Histologically, majority of the cases (54.83%) were diagnosed as chronic cholecystitis, followed by chronic cholecystitis with cholelithiasis (14.51%). Total 8 (12.91%) patients were found to have carcinoma of gallbladder with 50% of them belonging to grade I. Conclusion: Predominance of female gender in gallbladder ailment is clear from this study. Cholecystitis is the most common gallbladder disease. Microscopy varieties of histomorphological changes, including metaplasia and precursor changes are suggestive of high risk for gallbladder malignancy. Therefore, prompt detailed histopathological analysis of the cholecystectomy specimens will help to confirm the benign nature of the disease or to detect any precursors of malignancy. Mediscope 2022;9(1): 1-4