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Excelsior tissue processor

Manufactured by Thermo Fisher Scientific
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The Excelsior tissue processor is a laboratory equipment used for the automated processing of tissue samples. Its core function is to dehydrate, clear, and infiltrate tissue samples with paraffin wax, preparing them for subsequent embedding and sectioning.

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Lab products found in correlation

Ab290, by Cell Signaling Technology (1 mentions) Hm 325, by Thermo Fisher Scientific (1 mentions) Axioscan microscope slide scanner, by Zeiss (1 mentions) Shandon finesse 325 microtome, by Thermo Fisher Scientific (1 mentions) Histo clear 2, by Electron Microscopy Sciences (1 mentions) Paraformaldehyde (pfa), by Merck Group (1 mentions) Ab5398, by Merck Group (1 mentions) Sodium chloride (nacl), by Fresenius (1 mentions) Rm2065 microtome, by Leica (1 mentions) Microtome, by Leica (1 mentions) Anti v5, by Thermo Fisher Scientific (1 mentions) Anti chat, by Merck Group (1 mentions) Anti tau1, by Merck Group (1 mentions) Anti tdp 43, by Proteintech (1 mentions) Superfrost plus, by Thermo Fisher Scientific (1 mentions) Rnalater, by Thermo Fisher Scientific (1 mentions)

9 protocols using excelsior tissue processor

1

Immunostaining and Imaging Workflow

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Following transcardial perfusion in 1× PBS, left brain hemispheres were fixed in 4% paraformaldehyde overnight (4°C) and processed using an Excelsior tissue processor (Thermo Fisher Scientific). Samples were then embedded in paraffin and sectioned at a thickness of 3 μm (Thermo Fisher Scientific, HM325). Immunostaining was performed as previously described (10 (link)). Briefly, following antigen retrieval, histological sections were blocked and permeabilized (5% donkey serum + 0.1% Triton X-100). Primary antibodies, anti-GFP (Abcam, ab290) and anti-HA (Cell Signaling Technology, clone C29F4: 3724S), to visualize AAV-transgene expression, were incubated on slides overnight (4°C). Alexa fluorophore-coupled secondary antibodies 488 and 568 (Molecular Probes) diluted in blocking buffer (5% donkey serum) were incubated on slides at room temperature for 1 hour before 4′,6-diamidino-2-phenylindole staining for 10 min. Slides were mounted for confocal imaging using an AxioScan microscope slide scanner (Zeiss).
Morey N., Przybyla M., van der Hoven J., Ke Y.D., Delerue F., van Eersel J, & Ittner L.M. (2022). Treatment of epilepsy using a targeted p38γ kinase gene therapy. Science Advances, 8(48), eadd2577.
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2

Paraffin Embedding Protocol for Tissue Samples

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Samples were fixed in Bouin's solution and dehydrated in 96% alcohol at room temperature for 24 h. Next, the samples were exposed to a solution with a 1:1 ratio of xylene to alcohol for 15 min and purified in a series of 2 xylene solutions for 30 min each. The samples were then exposed to a solution with a 1:1 ratio of xylene to paraffin in a 60°C incubator for 15 min followed by exposure to a series of 2 paraffin solutions for 60 min each. After routine tissue processing (Shandon, Excelsior tissue processor, Thermo Fisher Scientific, Waltham, MA, USA), the tissues were embedded in paraffin.
Mete F., Tarhan H., Celik O., Akarken I., Vural K., Ekin R.G., Aydemir I, & Ilbey Y.O. (2016). Comparison of intraperitoneal and intratesticular ozone therapy for the treatment of testicular ischemia-reperfusion injury in rats. Asian Journal of Andrology, 19(1), 43-46.
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3

Tissue Sampling and Preparation Protocol

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After the completion of each experimental protocol, the brain, heart, lungs, kidneys, liver and pancreas were sampled and immersion-fixed in a solution containing 10% (w/v) neutral buffered formalin with gentle shaking. Sampling was performed by the same person at each experimental time point, randomly from the same region of each organ. After fixation, each tissue piece was processed and embedded into a paraffin block using the Excelsior tissue processor (Thermo Fisher Scientific). These paraffin blocks were trimmed on the Shandon Finesse 325 microtome (Thermo Fisher Scientific) to 5 μm sections. Sections were mounted on TruBond 380 adhesive slides and allowed to dry overnight at room temperature. All of the slides for the following tissue analysis were processed with deparaffinization and rehydration as previously described3 (link). In brief, the sections were deparaffinized in 2 changes of Histo-Clear II (64111–04, Electron Microscopy Sciences) for 10 min each. The slides were then transferred to 100% alcohol, for two changes, 10 min each, and then transferred once through 95%, 70% and 50% alcohol for 5 min each. The slides were then rinsed in water and washed in wash buffer (0.05% Tween-20 in 1× PBS) for 10 min.
Andrijevic D., Vrselja Z., Lysyy T., Zhang S., Skarica M., Spajic A., Dellal D., Thorn S.L., Duckrow R.B., Ma S., Duy P.Q., Isiktas A.U., Liang D., Li M., Kim S.K., Daniele S.G., Banu K., Perincheri S., Menon M.C., Huttner A., Sheth K.N., Gobeske K.T., Tietjen G.T., Zaveri H.P., Latham S.R., Sinusas A.J, & Sestan N. (2022). Cellular recovery after prolonged warm ischaemia of the whole body. Nature, 608(7922), 405-412.
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4

Developing Fetal Pancreatic Tissue

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Fourteen human fetal pancreatic specimens between 9 and 22 weeks of gestation (W9-W12, n = 5; W14-W22, n = 9) were collected from elective abortion tissue obtained by vacuum aspiration. “Weeks of gestation” used in this study is based on the last menstrual period (LMP), to convert to “weeks post conception” one need to subtract two weeks. This study was approved by the Medical Ethics Committee of the Leiden University Medical Center (protocol 08.087). Informed consent was obtained on the basis of the Declaration of Helsinki by the World Medical Association (WMA). All pancreata were fixed in 4% (w/v) paraformaldehyde (MERCK, Darmstadt, Germany) in PBS overnight at 4°C. Fixation was followed by dehydration in ethanol, xylene and paraffin embedding using standard procedures. Embedding was performed using a Shandon Excelsior tissue processor (Thermo Scientific, Altrincham, UK).
Roost M.S., van Iperen L., de Melo Bernardo A., Mummery C.L., Carlotti F., de Koning E.J, & Chuva de Sousa Lopes S.M. (2014). Lymphangiogenesis and angiogenesis during human fetal pancreas development. Vascular Cell, 6, 22.
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5

Archived Cardiac Tissue Collection

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Heart tissue was collected in the period 2003–2011 from deceased individuals autopsied at the Section of Forensic Pathology, Department of Forensic Medicine, Faculty of Health and Medical Sciences, University of Copenhagen, Denmark (Supplementary Table S1). The heart tissue was taken from deceased individuals with no or very little clinical evidence of tissue decomposition. After the autopsy, the heart tissue was fixed in 4% buffered formaldehyde (10% buffered formalin) for 48 h and embedded in paraffin using an Excelsior Tissue Processor (Thermo Fisher Scientific, Waltham, MA, USA), and archived at room temperature.
Dupont M.E., Christiansen S.N., Jacobsen S.B., Kampmann M.L., Olsen K.B., Tfelt-Hansen J., Banner J., Morling N, & Andersen J.D. (2023). DNA quality evaluation of formalin-fixed paraffin-embedded heart tissue for DNA methylation array analysis. Scientific Reports, 13, 2004.
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6

Immunohistochemistry of TRPV2 in Tissues

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Tissues were fixed overnight in 4% paraformaldehyde and further dehydrated with sequential 30%, 50% and 70% ethanol before final processing using Excelsior Tissue Processor according to manufacturer’s instructions (Thermo Scientific). Tissues were then embedded in paraffin and sectioned (5 μm) for H&E staining and immunohistochemistry as previously described [7 (link),18 (link)]. Antibodies were as follows: Anti-vanilloid receptor like protein 1 (TRPV2; Millipore, AB5398, 1:1000).
Naticchioni M., Karani R., Smith M.A., Onusko E., Robbins N., Jiang M., Radzyukevich T., Fulford L., Gao X., Apel R., Heiny J., Rubinstein J, & Koch S.E. (2015). Transient Receptor Potential Vanilloid 2 Regulates Myocardial Response to Exercise. PLoS ONE, 10(9), e0136901.
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7

Gonad Fixation and Sex Genotyping

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Gonads were isolated in cold 0.9% w/v NaCl (Fresenius Kabi), fixed in 4% paraformaldehyde (PFA) (MERCK) overnight (o/n) at 4°C, washed in phosphate-buffered saline (PBS) and transferred to a 70% ethanol solution followed by paraffin embedding using a Shandon Excelsior tissue processor (Thermo Scientific, Altrincham, UK). The material was sectioned (5 μm) using a RM2065 microtome (Leica Instruments GmbH, Wetzlar, Germany) onto StarFrost slides (Waldemar Knittel). Sex genotype was performed by PCR amplification of AMELX and AMELY, with fragments of 977 bp from X chromosome and 790 bp from Y chromosome. PCR conditions and primers were as previously described (Heeren et al., 2015 (link)).
Gomes Fernandes M., He N., Wang F., Van Iperen L., Eguizabal C., Matorras R., Roelen B.A, & Chuva De Sousa Lopes S.M. (2017). Human-specific subcellular compartmentalization of P-element induced wimpy testis-like (PIWIL) granules during germ cell development and spermatogenesis. Human Reproduction (Oxford, England), 33(2), 258-269.
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8

Embryo Collection and Spinal Cord Processing

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For the collection of embryos, C57Bl/6 females were bred with Hb9 V5-PFN1C71GWt/Tg males, using a timed mating protocol, where the time of plug check was considered to be E0.5. Pregnant females were euthanized by cervical dislocation at the appropriate timepoint. Embryos were harvested and immersion fixed in ice-cold 4% paraformaldehyde (PFA) for 90–120 min. Adult mice were anesthetized and transcardially perfused with ice-cold 1× phosphate-buffered saline (PBS). Brains were collected and fixed overnight in 4% PFA at 4°C.
The tissues were processed with an ethanol and xylene gradient in an Excelsior tissue processor (Thermo). Whole embryos were embedded in paraffin blocks. Spinal cords were cut into 2 mm segments and embedded into paraffin blocks with cervical to sacral sections arranged in a grid-like pattern. Samples were sectioned with a 5 μm thickness on a microtome (Leica). Sections were rehydrated and stained as previously described (Ittner et al., 2016 (link)). Antibodies used in this study were: anti-ChAT (Millipore), anti-profilin 1 (Abcam), anti-tau1 (Millipore), anti-TDP-43 (Proteintech) and anti-V5 (Invitrogen, Carlsbad, CA, USA).
Brettle M., Stefen H., Djordjevic A., Fok S.Y., Chan J.W., van Hummel A., van der Hoven J., Przybyla M., Volkerling A., Ke Y.D., Delerue F., Ittner L.M, & Fath T. (2019). Developmental Expression of Mutant PFN1 in Motor Neurons Impacts Neuronal Growth and Motor Performance of Young and Adult Mice. Frontiers in Molecular Neuroscience, 12, 231.
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9

Tissue Preparation for Histology

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Tissues were dissected, weighed (liver, spleen, and kidney), and placed in 10% neutral buffered formalin or RNAlater (Ambion). For histology, tissues were processed overnight using an Excelsior tissue processor (Thermo Fisher Scientific). Sections were embedded in paraffin wax prior to 4-μm sections cut and mounted onto slides (Superfrost Plus, Thermo Fisher Scientific). Slides were dried overnight at 37°C before 60°C for 25 min. Sections were stained with H&E or immunohistochemistry was performed by R(D)SVS pathology department.
Sauter K.A., Waddell L.A., Lisowski Z.M., Young R., Lefevre L., Davis G.M., Clohisey S.M., McCulloch M., Magowan E., Mabbott N.A., Summers K.M, & Hume D.A. (2016). Macrophage colony-stimulating factor (CSF1) controls monocyte production and maturation and the steady-state size of the liver in pigs. American Journal of Physiology - Gastrointestinal and Liver Physiology, 311(3), G533-G547.
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