title: Optimization and validation of cryostat temperature conditions for trans-reflectance mode FTIR microspectroscopic imaging of biological tissues.
abstract: In Fourier transform infrared (FTIR) microspectrocopy, the tissue preparation method is crucial, especially how the tissue is cryo-sectioned prior to the imaging requires special consideration. Having a temperature difference between the cutting blade and the specimen holder of the cryostat greatly affects the quality of the sections. Therefore, we have developed an optimal protocol for cryo-sectioning of biological tissues by varying the temperature of both the cutting blade and the specimen holder. Using this protocol, we successfully cryo-sectioned four different difficult-to-section tissues including white adipose tissue (WAT), brown adipose tissue (BAT), lung, and liver. The optimal temperatures that required to be maintained at the cutting blade and the specimen holder for the cryo-sectioning of WAT, BAT, lung, and liver are (-25, -20 °C), (-25, -20 °C), (-17, -13 °C) and (-15, -5 °C), respectively. The optimized protocol developed in this study produced high quality cryo-sections with sample thickness of 8-10 μm, as well as high quality trans-reflectance mode FTIR microspectroscopic images for the tissue sections. •Use of cryostat technique to make thin sections of biological samples for FTIR microspectroscopy imaging.•Optimized cryostat temperature conditions by varying the temperatures at the cutting blade and specimen holder to obtain high quality sections of difficult-to-handle tissues.•FTIR imaging is used to obtain chemical information from cryo-sectioned samples with no interference of the conventional paraffin-embedding agent and chemicals.
authors.affiliation: Fiber and Biopolymer Research Institute, Department of Plant and Soil Science, Texas Tech University, Lubbock, TX 79403, USA.
authors.forename: Sumedha
authors.initials: S
authors.lastname: Liyanage
authors.affiliation: Fiber and Biopolymer Research Institute, Department of Plant and Soil Science, Texas Tech University, Lubbock, TX 79403, USA.
authors.forename: Rohan S
authors.initials: RS
authors.lastname: Dassanayake
authors.affiliation: Fiber and Biopolymer Research Institute, Department of Plant and Soil Science, Texas Tech University, Lubbock, TX 79403, USA; Department of Nutritional Sciences, Texas Tech University, Lubbock, TX 79409, USA.
authors.forename: Amal
authors.initials: A
authors.lastname: Bouyanfif
authors.affiliation: Fiber and Biopolymer Research Institute, Department of Plant and Soil Science, Texas Tech University, Lubbock, TX 79403, USA.
authors.forename: Erandathi
authors.initials: E
authors.lastname: Rajakaruna
authors.affiliation: Department of Nutritional Sciences, Texas Tech University, Lubbock, TX 79409, USA; Obesity Research Cluster, Texas Tech University, Lubbock, TX 79409, USA.
authors.forename: Latha
authors.initials: L
authors.lastname: Ramalingam
authors.affiliation: Department of Nutritional Sciences, Texas Tech University, Lubbock, TX 79409, USA; Obesity Research Cluster, Texas Tech University, Lubbock, TX 79409, USA.
authors.forename: Naima
authors.initials: N
authors.lastname: Moustaid-Moussa
authors.affiliation: Fiber and Biopolymer Research Institute, Department of Plant and Soil Science, Texas Tech University, Lubbock, TX 79403, USA.
authors.forename: Noureddine
authors.initials: N
authors.lastname: Abidi
keywords: Cryo-sectioning and FTIR microspectroscopic imaging of biological tissues; Cryostat; FTIR; Microspectroscopic imaging; Tissue sectioning