Cx31 optical microscope

Coronary tissues were fixed in 10% formalin (NanJing JianCheng Bioengineering Institute, Nanjing, China) for 24 h, followed by dehydration with ethanol (Sigma-Aldrich, St. Louis, Missouri, USA), clearance with xylene, dewaxing, and embedment in paraffin. After that, the paraffin-embedded block was dewaxed and stained with hematoxylin staining solution. Next, the tissues were differentiated by 1% acidic alcohol, immersed in eosin solution, and dehydrated in ethanol. After clearance with xylene twice (10 min), the tissues were covered with Acacia senegal and observed under a CX-31 optical microscope (Olympus, Tokyo, Japan). An LM1235 ultra-thin semi-automatic slicer (Leica, Wetzlar, Germany) was utilized for sectioning while a KD-BM machine for tissue embedment (Kedee, Zhejiang, China).

After ex vivo studies, each tissue was fixed in buffered 10% formalin and embedded in paraffin wax. Four-micrometer-thick sections of the tissue were stained with hematoxylin-eosin and were placed on microscope slides. The tissue sections were analyzed with an Olympus CX31 optical microscope and images were processed with OLYMPUS analysis getIT software (Version 5-1, Shinjuku-ku, Tokyo, Japan).

Optical micrographs of MAP and MCMAP were obtained using an Olympus CX 31 optical microscope attached to a computer system. Samples were placed on glass slides, and a few drops of ethanol were added to disperse the powders. A cover glass was then placed on top of each suspension, and the micrographs were captured using a computer system.

The optical observations of the CCS microgranules were carried out using an Olympus CX31 optical microscope (Cebu, Philippines) under 100-time magnification. The samples were prepared by dyeing cationic starch derivative microgranules with an 0.005 M standard iodine solution. The photographs of the CCS slurry were taken using an Olympus camera.

Flower buds with different lengths were fixed in formalin-aceto-alcohol (FAA), dehydrated in an ethanol series, embedded in paraffin, sectioned into 3–5 μm transverse slices using a microtome and stained with 1% toluidine blue as described by Lou et al. [40 (link)]. Then the anther transverse sections were observed with an Olympus CX31 optical microscope (Olympus Japan Co., Tokyo, Japan) and photographed with a Nikon 550D camera (Canon, Tokyo, Japan).

Histochemical tests were carried out in Laboratório de Anatomia Vegetal of Departamento de Biologia from Universidade Federal de Lavras, Brazil, according to the methodology proposed by Figueiredo et al. ([2007 ]). The tests were applied on 3 leaves/individual of D. brasiliensis, being 4 individuals/altitude level (1900 and 2100 m). The cross sections were carried out from approximately 2 cm² fragments, taken from the region that contains the central vein with the help of an LPC bench microtome.
For the detection of total lipids, the reagent Sudan IV was used; for the detection of the essential oils, the reagent NADI was used, and for the verification of the phenolic compounds, ferric chloride was used. The sections were mounted on slides and coverslips with 50% glycerin, and photographed in an Olympus CX 31 optical microscope, coupled to a digital camera.