Cryovial

Manufactured by Thermo Fisher Scientific

Sourced in United States, Denmark, Germany, United Kingdom

Cryovials are laboratory storage containers designed for the long-term preservation of samples at ultra-low temperatures, typically in cryogenic freezers or liquid nitrogen. They are made of high-quality materials that are resistant to the extreme cold conditions required for sample storage. Cryovials come in various sizes and feature screw-top or snap-top lids to maintain sample integrity.

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Lab products found in correlation

50 protocols using cryovial

Cryopreservation and Thawing of ADSCs

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Approximately 1 × 10⁶ ADSCs (P3) were resuspended in 1 ml CPAs and transferred into cryovials (Thermo Fisher, Waltham, MA, USA). The cryovials were frozen in a Nalgene® Mr. Frosty freezing container (Thermo Fisher, Waltham, MA, USA) at a cooling rate of 1 °C/min to − 80 °C, stored overnight, and transferred into liquid nitrogen for 30 days for storage. For thawing, the cryovials were placed in a water bath at 37 °C under gentle shaking until the ice was completely melted. The thawed ADSCs were rinsed with 10 ml PBS by centrifugation at 1500 rpm for 5 min (2 times) using a Microfuge 20/20R centrifuge (Beckman, USA) and resuspended in 5 ml Dulbecco’s modified Eagle’s medium (DMEM)/F12 containing 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin for further assessment.

Zhang T.Y., Tan P.C., Xie Y., Zhang X.J., Zhang P.Q., Gao Y.M., Zhou S.B, & Li Q.F. (2020). The combination of trehalose and glycerol: an effective and non-toxic recipe for cryopreservation of human adipose-derived stem cells. Stem Cell Research & Therapy, 11, 460.

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Cryopreservation of Human Adipose-Derived Stem Cells

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To cryopreserve hADSCs, the cells were seeded in 6-well plate at a density of 1 × 10⁵ cells/well in 1 ml medium. After overnight incubation, the cells were further incubated in medium with GNPs, fTre, or nTre for 24 h. Cells incubated in medium without any further supplement (i.e., no pre-treatment or NPT) were also prepared in the same way for control. The cells were then washed with 1x PBS, detached using trypsin/EDTA, and collected by centrifugation at 188 g for 5 min. The collected cells were then washed with 1x PBS and re-suspended in 200 μl hADSC culture medium containing 200 mM free trehalose. The cell suspension was transferred into a 1 ml cryovial (Fisher scientific, Pittsburgh, PA, USA) and the cryovial was hermetically sealed before putting in a Thermo Nalgene freezing container that cools/freezes the sample at approximately -1 °C/min to −80 °C in a −80 °C freezer. On the second day, the cryovials were transferred into liquid nitrogen tank for storage. For cryopreservation with DMSO, the detached hADSCs were re-suspended in 200 μl cell culture medium with 10% v/v DMSO in 1 ml cryovial and cryopreserved in the same way using the Thermo Nalgene freezing container. After 1 day storage, the cryovials were removed from the liquid nitrogen tank and thawed in 37 °C water bath. The cells were then washed with 10 ml medium for further analysis.

Rao W., Huang H., Wang H., Zhao S., Dumbleton J., Zhao G, & He X. (2015). Nanoparticle-Mediated Intracellular Delivery Enables Cryopreservation of Human Adipose-Derived Stem Cells Using Trehalose as the Sole Cryoprotectant. ACS applied materials & interfaces, 7(8), 5017-5028.

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Isolation and Cryopreservation of Blood Cells

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Blood was centrifuged in vacutainers used for collection at 2000 rpm for 10 min. Plasma was aspirated from the top and frozen at −80 °C in 1-mL aliquots in cryovials (Thermo Fisher Scientific, cat. #375418) using a freeze controller (Bel-Art Products, cat. #F18844-0000) pre-chilled to −4 °C according to the manufacturer’s instructions. The remaining blood was diluted 1:1 in PBS without calcium or magnesium, layered over 15 mL of Ficoll-Paque (GE Healthcare, cat. #17-1440-03) in an Accuspin tube (Sigma-Aldrich, cat. #A2055), and centrifuged at 2000 rpm for 20 min at 21 °C with acceleration at five and break at zero. The buffy coat leukocyte layer was collected and washed twice in 50 mL of PBS without calcium or magnesium by centrifuging at 2000 rpm for 10 min. Cells were counted, washed again, and resuspended in the Recovery Cell Culture Freezing Medium (Thermo Fisher Scientific, cat. #12648010) at 3.5–10 × 10⁶ cells/mL in 1-mL aliquots, transferred to a freeze controller (Bel-Art Products, cat. #F18844-0000) pre-chilled to −4 °C according to the manufacturer’s instructions, stored at −80 °C for 1–7 days, and then transferred to liquid nitrogen for storage.

Rubin S.J., Bai L., Haileselassie Y., Garay G., Yun C., Becker L., Streett S.E., Sinha S.R, & Habtezion A. (2019). Mass cytometry reveals systemic and local immune signatures that distinguish inflammatory bowel diseases. Nature Communications, 10, 2686.

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Cryopreservation of iPSC-derived Cardiomyocytes

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iPSC‐derived cardiomyocytes were collected from the bioreactor on Day 14, washed with phosphate buffered saline (PBS)^−/− (ThermoFisherScientific) once and resuspended in Liberase solution (50 μg/ml; Roche) and incubated at 37°C for 30 min. The aggregates were triturated every 15 min using 10 ml serological pipettes (Corning). Following dissociation, an equal volume of growth medium (RPMI‐1640 + B27 with insulin) was added to the cell suspension to dilute the Liberase. The cells were centrifuged at 200g for 3 min at room temperature. Following centrifugation, the supernatant was removed and discarded. TrypLE solution was added to the cells and incubated for 7–8 min at 37°C. The enzyme was subsequently diluted by adding equal volume of growth medium. Viable cell counts were performed with the NC‐200 and the cells centrifuged at 200g for 3 min at 4°C. The supernatant was removed, discarded and the cell pellet was resuspended in CryoStor CS10 (BioLife Solutions) supplemented with 10 μM Y‐27632 at 5 × 10⁶ cells/ml. Cryovials (ThermoFisherScientific) were filled at 1.0 ml and cryopreserved using a Controlled Rate Freezer (ThermoFisherScientific).

Shafa M., Panchalingam K.M., Walsh T., Richardson T, & Baghbaderani B.A. (2019). Computational fluid dynamics modeling, a novel, and effective approach for developing scalable cell therapy manufacturing processes. Biotechnology and Bioengineering, 116(12), 3228-3241.

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Anaerobic Bacterial Isolation from Biopsies

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All collected biopsies were placed in cryovials (Thermo Fisher Scientific Inc. cat# 368632, USA) filled with anaerobic media (Anaerobic systems, Cat# AS-916, USA) and processed fresh in the 30 minutes following collection. Following immediate and thorough homogenization with a homogenizing pestle (Thermo Fisher Scientific Inc. Cat# K749521-1590, USA) in an anaerobic chamber and under sterile conditions, samples were plated on non-selective Tryptic Soy Agar containing 5% sheep blood (Anaerobic systems, Cat# AS-542, USA) for bacterial growth. The plates were kept under anaerobic conditions (90% N₂, 5% CO₂, 5% H₂) at 37°C for 2 days. Colonies cultured for 48 hours were expanded in Tryptic Soy Broth + 5% defibrinated sheep blood after identification, then transferred to 50% skim milk stock (BD Difco Skim Milk, Cat# 232100, USA), and preserved at −80°C until analyzed.

Kordahi M.C., Stanaway I.B., Avril M., Chac D., Blanc M.P., Ross B., Diener C., Jain S., McCleary P., Parker A., Friedman V., Huang J., Burke W., Gibbons S.M., Willis A.D., Darveau R.P., Grady W.M., Ko C.W, & DePaolo R.W. (2021). Genomic and functional characterization of a mucosal symbiont involved in early-stage colorectal cancer. Cell host & microbe, 29(10), 1589-1598.e6.

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Quantification of Intracellular Pyruvate and Acetyl-CoA

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Intracellular pyruvate (cPyr) and acetyl‐CoA (cAcCoA) concentrations were quantified with assay kits (#ab65342 and #ab87546, Abcam, USA). The cells harvested from the bioreactor (≈2·106 cells) were washed with PBS (1000g, 3 min) and the cell pellets were snap‐frozen and stored in liquid nitrogen (1.8 ml cryovials, Thermo Scientific, Germany). For the quantification of intracellular metabolites, the snap‐frozen cell pellets were resuspended in assay buffer (containing surfactant, 4∘C) and incubated for 5 min on ice with intermediate vortexing. Then, the suspension was centrifuged at 21100g for 5 min at 4∘C to obtain debris‐free lysate (i.e., supernatant). Deproteinization was carried out using a 10 kDa spin filter (Amicon Ultra‐0.5 mL Centrifugal Filter, Merck, Germany) with centrifugation at 14000g for 10 min at 4∘C. Subsequently, the assays were carried out according to the manufacturer's protocol.

Möller J., Bhat K., Guhl L., Pörtner R., Jandt U, & Zeng A. (2020). Regulation of pyruvate dehydrogenase complex related to lactate switch in CHO cells. Engineering in Life Sciences, 21(3-4), 100-114.

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Biomarker Profiling in Alzheimer's and NPH

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This study involved 36-AD, 32-iNPH, and 12-healthy age-matched subjects (Table 1 and S1 Table), enrolled by Fondazione Ca’ Granda, IRCCS Ospedale Maggiore Policlinico, Milan, Italy.
All participants gave their informed consent to the study, including medical history, physical and neurological examination, neurocognitive evaluation (Mini-Mental State Examination), computed tomography or MRI scan, and screening laboratory tests consisting in the assessment of levels of tau, phospho-tau (p-tau), and amyloid-β (Aβ) proteins by ELISA (Innogenetics). AD patients fulfilled the NINCDS-ADRDA criteria [20 (link)] and those proposed by McKhann et al. [21 (link)]. The iNPH subjects were diagnosed according to International Guidelines published in 2005 [22 (link)]. Control subjects were likewise examined to exclude the presence of neurological and cognitive disorders.
CSF samples were drawn in polypropylene tubes after lumbar puncture at the L4/L5 or L3/L4 interspace, centrifuged at 4°C and stored at ≤ −80°C until analysis.
Blood samples were collected in vacutainer tubes and placed at 4°C for 15 min until clotted. Samples were centrifuged for 30 min at 3000 rpm at 4°C and sera transferred into 1.8 mL cryovials (Thermo Fisher Scientific) and stored at −80°C.

Fania C., Arosio B., Capitanio D., Torretta E., Gussago C., Ferri E., Mari D, & Gelfi C. (2017). Protein signature in cerebrospinal fluid and serum of Alzheimer’s disease patients: The case of apolipoprotein A-1 proteoforms. PLoS ONE, 12(6), e0179280.

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Cryopreservation of Testicular Tissue

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Upon surgical removal, pairs of whole testes were put in containers on ice for 1 to 2 hours of transportation. Then the tunica were removed and testicular tissues were cut into small pieces (~500 mg to 1g each). 90% of tissues were directly transferred into cryovials (Corning) containing 1.5ml freezing medium (75% DMEM medium (Life Technologies) +10% DMSO (Sigma-Aldrich cat # D8779) + 15% fetal bovine serum (FBS) (Gibco)). The cryovials are placed in the isopropanol chamber (Thermo Fisher Scientific) and stored at −80°C overnight. Then the cryovials were transferred to liquid nitrogen for long-term storage.

Nie X., Munyoki S.K., Sukhwani M., Schmid N., Missel A., Emery B.R., Stukenborg J.B., Mayerhofer A., Orwig K.E., Aston K.I., Hotaling J.M., Cairns B.R, & Guo J. (2022). Single-Cell Analysis of Human Testis Aging and Correlation with Elevated Body Mass Index. Developmental cell, 57(9), 1160-1176.e5.

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Isolation and Cryopreservation of PBMCs

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A total volume of 20 mL blood was collected from PCa patients at the time of enrollment, which was subsequently used for HLA Class I typing and isolation of peripheral blood mononuclear cells (PBMCs). PBMCs were isolated from blood samples by Ficoll (Biochrom, Holliston, MA, USA) gradient separation at RT, washed twice with PBS, and counted in a Neubauer chamber (Poly-optik GmbH, Bad Blankenburg, Germany). Viability was always >95%. Cells were resuspended in RPMI + 40% FCS (all from Thermofisher, Waltham, MA, USA) at a concentration of 10 × 10⁶ /mL, and half of the volume of the same medium containing 20% DMSO (Applichem GmbH, Darmstadt, Germany) was added quickly at room temperature. After thorough mixing, DMSO was allowed to equilibrate through the cell membrane for 5 min before a second aliquot of the medium was added to bring the final concentration of DMSO to 10%. Next, 1 mL of the cell suspension was immediately transferred into cryovials (Thermofisher, Waltham, MA, USA) and placed in boxes at −80 °C overnight and then stored in liquid nitrogen until use.

Goulielmaki M., Stokidis S., Anagnostou T., Voutsas I.F., Gritzapis A.D., Baxevanis C.N, & Fortis S.P. (2023). Frequencies of an Immunogenic HER-2/neu Epitope of CD8+ T Lymphocytes Predict Favorable Clinical Outcomes in Prostate Cancer. International Journal of Molecular Sciences, 24(6), 5954.

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Whole blood sample processing and storage

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Whole blood samples were collected using BD Vacutainer® blood collection tubes (BD Life Science, Franklin Lakes, NJ, USA). Within 2 h of collection, the samples were centrifuged at 1100× g for 10 min and filtered using 0.8 μm filters prior to being aliquoted into cryovials (Thermo Fisher Scientific, Waltham, MA, USA). All the aliquoted samples were stored at −80 °C.

Wang L., Yekula A., Muralidharan K., Small J.L., Rosh Z.S., Kang K.M., Carter B.S, & Balaj L. (2020). Novel Gene Fusions in Glioblastoma Tumor Tissue and Matched Patient Plasma. Cancers, 12(5), 1219.

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