بررسی مقایسه ‏ای هیستومورفومتری بافت روده باریک با سه روش کلارک-بسته، فرمالین-باز و فرمالین-بسته در جوجه‏های گوشتی

محورهای موضوعی : آسیب شناسی درمانگاهی دامپزشکی

عارف محمودتبار ¹ , محمد امیر کریمی ترشیزی ²

1 - گروه پرورش و مدیریت طیور، دانشکده کشاورزی، دانشگاه تربیت مدرس، تهران، ایران
2 - گروه پرورش و مدیریت طیور- دانشکده کشاورزی دانشگاه تربیت مدرس- تهران- ایران

تاریخ دریافت : 1396/10/15 تاریخ پذیرش : 1397/09/14 تاریخ انتشار : 1397/10/01

کلید واژه: فرمالین, روده باریک, محلول کلارک, هیستومرفومتری, پایدار‌سازی بافت,

چکیده مقاله :

پایدار نمودن بافت یک مرحله بسیار حیاتی در آماده سازی بافت جهت مطالعات بافت شناسی به شمار می رود و هرگونه اشتباه در این مرحله غیرقابل جبران خواهد بود. هدف از این مطالعه مقایسه تاثیر سه روش تهیه مقاطع بافتی کلارک-بسته، فرمالین-باز و فرمالین-بسته بر هیستومورفومتری مخاط روده باریک در جوجه های گوشتی بود. در این بررسی 30 قطعه جوجه گوشتی سویهراس 308 نر در سن 10 روزگی مورد استفاده قرار گرفت. نمونه های روده باریک بعد از شستشو، بلافاصله با محلول سالین- فرمالین 10 درصد به‌شکل بسته و یا بازشده و محلول کلارک به‌صورت دو‌طرف بسته پایدار شدند. پس از پایداری طبق روش استاندارد بافت‌شناسی، مقاطع میکروسکوپی به ضخامت 5 میکرومتر تهیه و با ائوزین-هماتوکسیلین و آلسین‌بلو رنگ‌آمیزی شدند. در هر مقطع ارتفاع پرز، ضخامت پرز، عمق کریپت و تعداد سلول های جامی با خط‌کش چشمی اندازه‌گیری و شاخص سطح پرز نیز محاسبه گردید. داده ها‌ در قالب طرح کاملاً تصادفی با آزمون دانکن مورد مقایسه قرار گرفتند. نتایج نشان داد که روش تهیه و پایداری نمونه‌های روده باریک بر شاخص های کمی بافت شناسی تاثیر معنی داری داشت (01/0>p). روش فرمالین-بسته بلند ترین ارتفاع پرز و بیشترین عمق کریپت و روش کلارک-بسته، کمترین مقادیر را نشان دادند (01/0>p). همچنین روش کلارک-بسته و روش های فرمالین-بسته و باز به ترتیب بیشترین ضخامت پرز را در هر سه قسمت روده کوچک نشان دادند (01/0>p) و بیشترین مقدار برای سطح پرز نیز در روش فرمالین-بسته مشاهده شد (01/0>p). با توجه به سادگی روش کار، قیمت مناسب، حفظ بهتر مخاط روده و عدم ایجاد تغییرات شدید در بافت، روش فرمالین-بسته به عنوان روش برگزیده معرفی می شود.

چکیده انگلیسی:

Tissue fixation is a vital step in preparation of samples for histological studies and mistakes at this stage could lead to irreversible damage. The aim of this study was to compare the effect of three sample preparation-fixation methods on histomorphometic indices of small intestine of broilers. Thirty 10 day old broilers of Ross 308 strain were used in the study. Segments of small intestine were washed and prepared as opened or closed sections. The closed sections were fixed in 10% saline-formalin. The opened samples were cut at the mesenteric line and were fixed by 10% saline-formalin. In Clarke’s fixation the ends of samples were sealed by string and Clarke’s solution was injected into the lumen. Fixed samples were subjected to paraffin embedding and 5 µm sections were cut using rotary microtome and sections were stained by Eosin-Hematoxylin and Alcian blue. In each section the villus height, villus thickness, crypt depth and goblets’ density were measured under light microscope and the villus surface index was also calculated. Data were subjected to ANOVA in a completely randomized design with three treatments and 30 observations. The results indicated that method of preparation-fixation had a significant effect on morphometric indices (p<0.01). The closed-formalin’s samples had the greatest villus height and crypt depth values in all three intestinal segments, while the lowest values were observed in closed-Clarke’s method (p<0.01). Closed-Clarke’s samples had the thickest villus in all three parts of small intestine followed by closed-formalin’s and opened-formalin’s samples (p<0.01). Villus surface area was highest in closed-formalin fixed samples (p<0.01). Considering simplicity of the method, low cost,  better mucosal protection and lack of severe tissue changes observed in closed-formalin’s method, this method was considered as a method of choice in histomorphometric studies of poultry small intestinal mucosa.

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