Microm rm55 rotary microtome

Internal organs from half of the mice in each treatment group were collected at necropsy and fixed in 10% neutral buffered formalin for 3 days and kept in 70% EtOH and 4°C thereafter. The tissue processing and hematoxylin and eosin (H&E) staining were done by ARUP Laboratories (Salt Lake City, Utah). Briefly, the tissues were processed using the Tissue-Tek VIP 6 Vacuum Infiltration Processor (Sakura Finetek, CA, USA) and embedded in molten paraffin wax using the Tissue-Tek Embedding Center. The tissues were then sliced on Leica Microm RM55 Rotary Microtome (IL, USA), and placed on glass slides. The slides were stained with H&E using the Tissue-Tek Prisma Automated Slide Stainer (Sakura Finetek, CA, USA). Slides were scanned at 20X by Aperio digital pathology slide scanner (Leica Biosystems, IL, USA) and analyzed via Image Scope software (Leica Biosystems, IL, USA).

The dissected organs during necropsy including lung, liver, brain, heart, intestine and genitourinary organs, were fixed in formalin for three days and then transferred to an ethanol solution (70%). Tissue samples were submitted in tissue embedding cassettes (Simport, Canada), processed by Tissue-Tek VIP 6 Vacuum Infiltration Processor (Sakura Finetek, CA, USA) at ARUP Laboratories (Salt Lake City, Utah). The sections were obtained from the prepared tissue blocks using Leica Microm RM55 Rotary Microtome (IL, USA) and placed on glass slides, later stained with Hematoxylin and Eosin on Tissue-Tek Prisma Automated Slide Stainer (Sakura Finetek, CA, USA). Slides were scanned with Aperio digital pathology slide scanner (Leica Biosystems, IL, USA) and analyzed with Image Scope software (Leica Biosystems, IL, USA).