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Formal fixx

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Formal-Fixx is a laboratory equipment product designed for the fixation of biological samples. It provides a standardized approach to preserving the structural integrity of cellular and tissue samples for further analysis and examination.

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Lab products found in correlation

Fluoro max g100, by Thermo Fisher Scientific (1 mentions) Rf5000 fluorescent spectrophotometer, by Shimadzu (1 mentions) Lsm700 upright confocal microscope, by Zeiss (1 mentions) Alexa fluor 488 phalloidin, by Thermo Fisher Scientific (1 mentions) Triton x 100, by Merck Group (1 mentions) Bovine serum albumin (bsa), by Merck Group (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions) Xylene, by Merck Group (1 mentions) Kainic acid ka, by Bio-Techne (1 mentions) Dpx mounting medium, by Merck Group (1 mentions) Valproic acid vpa, by Merck Group (1 mentions) Carbamazepine cbz, by Merck Group (1 mentions) Hoeschst 33342, by Thermo Fisher Scientific (1 mentions) Sodium hydroxide (naoh), by Thermo Fisher Scientific (1 mentions) Potassium permanganate kmno4, by Merck Group (1 mentions) Ketamine ketaset, by Zoetis (1 mentions) Y 27632 dihydrochloride, by Bio-Techne (1 mentions) Sodium chloride (nacl), by Thermo Fisher Scientific (1 mentions) Hepes, by Thermo Fisher Scientific (1 mentions) Dimethyl sulfoxide (dmso), by Thermo Fisher Scientific (1 mentions)

Spelling variants of this product

Zinc Formal-Fixx (10 citations) Shandon Formal-Fixx (6 citations) Shandon Zinc Formal-Fixx (3 citations) Formal-Fixx formalin solution (2 citations) Shandon Formal-Fixx 10% neutral buffered formalin (2 citations)

10 protocols using formal fixx

1

Quantifying Exosome Uptake by Cells

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To quantify the amount of exosomes taken up by cells, exosomes were stained with a lipophilic fluorescent dye, DIL as described previously (29 ). Then, DIL-labeled exosomes, or fluorescently-labeled liposomes, or polystyrene nanoparticles (NPs, Fluoro-Max G100, Thermo Fisher Scientific), were added in equal numbers (~108 particles/well) and incubated with 3LL-M27 cells at 37°C and 5% CO2 for various times. After each time point, the media was removed and cells were washed 3x with PBS and fixed by incubating with Formal-Fixx (Thermo Fisher Scientific), and examined by confocal microscopy or using a Shimadzu RF5000 fluorescent spectrophotometer. In case of exoPTX or Taxol, drugs were added in equimolar amounts to the MDCKWT, or MDCKMDR1 cells and incubated for 72h. The cell suspension was then lysed and analyzed for PTX content by HPLC as described above.
Kim M.S., Haney M.J., Zhao Y., Mahajan V., Deygen I., Klyachko N.L., Inskoe E., Piroyan A., Sokolsky M., Okolie O., Hingtgen S.D., Kabanov A.V, & Batrakova E.V. (2015). Development of Exosome-encapsulated Paclitaxel to Overcome MDR in Cancer cells. Nanomedicine : nanotechnology, biology, and medicine, 12(3), 655-664.
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2

PINK1 Knockout Brain Harvesting

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Brains were rapidly harvested in accordance with the IACUC protocols at the University of Nebraska Medical Center. Six brains were taken from 9 month old animals from both the PINK1 knockout (PINK1 KO) and Long Evans Hooded (LEH) control groups. Brains were placed in Formal Fixx (Thermo Scientific, Rockford, IL) overnight (~15 hours). Brains were immersed in 30% sucrose in 0.1 M PBS overnight at 4° C. Brains were place in 70% ethanol and shipped on dry ice to the Stereology Resource Center, Inc. for stereology.
Villeneuve L.M., Purnell P.R., Boska M.D, & Fox H.S. (2014). Early expression of Parkinson’s disease-related mitochondrial abnormalities in PINK1 knockout rats. Molecular neurobiology, 53(1), 171-186.
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3

Mitochondrial Visualization and Analysis

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Cells were fixed with Formal-Fixx (Thermo Scientific) and permeabilized with 0.4% (v/v) Triton X-100 for 5 minutes. Images were acquired using a Zeiss LSM 700 Upright confocal microscope (Carl Zeiss AG) under non-saturating exposure conditions. For each condition multiple cells were observed and imaged. For the uniformity of the represented images, contrast and brightness were adjusted to eliminate undesirable background signal. Neither of these manipulations was affecting the mitochondrial shape. Image processing was performed with the Fiji software (http://imagej.nih.gov/ij; version 1.47b).
Moullan N., Mouchiroud L., Wang X., Ryu D., Williams E.G., Mottis A., Jovaisaite V., Frochaux M.V., Quiros P.M., Deplancke B., Houtkooper R.H, & Auwerx J. (2015). Tetracyclines disturb mitochondrial function across eukaryotic models: a call for caution in biomedical research. Cell reports, 10(10), 1681-1691.
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4

Chewing Betel Nut and Chocolate: Buccal Cell Analysis

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Betel nut quid was prepared using one (1) white areca nut (husk included), one (1) betel leaf, and approximately a 1/2 teaspoon of slaked lime (all locally sourced in Honolulu, Hawaii). Buccal cells from healthy male volunteers were collected prior to the exposure to BQ or other substances. For nine minutes, a volunteer chewed either BQ or a 72% cacao chocolate bar as a control for an optically active compound. Buccal cells were also collected 24 hours following exposure to all substances. Cells were collected by gently scraping a soft-bristle toothbrush up and down five times on the buccal cell region. The brushes were inserted into 15 ml of PBS inside a 50 ml conical tube, covered with parafilm, and vortexed for 15 seconds to loosen the cells adhered to the bristles and suspend them in the PBS. The tubes were then centrifuged at 1000 × g for 5 min. The cells were then fixed and frozen according to a modified protocol from the BD Biosciences website (www.bdbiosciences.com/us/resources/s/cytokinesfca). Briefly, cells were mixed with 1X Formal-Fixx (Thermo Fisher Scientific, Waltham, MA) for 20 mins at 4°C then centrifuged at 500 × g for 5 mins, resuspended with cryoprotectant mixture (500 μl 90% FBS/10% DMSO), and stored at −80°C until analysis by flow cytometry and microscopy.
Biggs L.A., Franke A.A, & Farrar C.E. (2020). Altered Fluorescence of Buccal Cells as a Candidate Biomarker for Areca Nut Chewing. Substance use & misuse, 55(9), 1450-1456.
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5

Lung Tissue Histological Analysis

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Lung tissue was collected on day 6 after adoptive cell transfer and OVA treatment and stored in buffered formalin (Formal-Fixx, Thermo Fisher Scientific) as per manufacturer’s instructions. Then the samples were transferred to 70% ethanol, and submitted to HistoWiz (https://home.histowiz.com) for tissue sectioning, Hematoxylin and Eosin (H&E) staining and imaging.
Canaria D.A., Clare M.G., Yan B., Campbell C.B., Ismaio Z.A., Anderson N.L., Park S., Dent A.L., Kazemian M, & Olson M.R. (2022). IL-1β promotes IL-9-producing Th cell differentiation in IL-2-limiting conditions through the inhibition of BCL6. Frontiers in Immunology, 13, 1032618.
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6

Fluorescent Labeling of Actin and Nuclei

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After being fixed in Formal-Fixx (ThermoFisher Scientific) for 10 min, cells were washed and blocked in phosphate-buffered solution (PBS) containing 3% bovine serum albumin (Sigma, Buchs, Switzerland) and 0.1% Triton X-100 (Sigma) for 10 min. Cells were then labeled with Alexa FluorTM 488 Phalloidin (Invitrogen, ThermoFisher Scientific) or Phalloidin-TRITC (Bio-Techne, Zug, Switzerland) at room temperature for 1 h. Cells were washed in PBS containing 0.1% Triton X-100 and co-stained with 4′,6-diamidino-2-phenylindole (DAPI; Sigma) at the last washing step before being mounted in Vectashield Medium (Adipogen, Fuellinsdorf, Switzerland). Images were acquired on an Olympus BX-51 (Olympus Life Sciences Solution, Tokyo, Japan) equipped with the fluorescent filters U-MWIBA3 for Alexa Fluor 488, U-MWIGA3 for TRITC, and U-MNUA2 for DAPI.
Asparuhova M.B., Riedwyl D., Aizawa R., Raabe C., Couso-Queiruga E, & Chappuis V. (2023). Local Concentrations of TGF-β1 and IGF-1 Appear Determinant in Regulating Bone Regeneration in Human Postextraction Tooth Sockets. International Journal of Molecular Sciences, 24(9), 8239.
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7

Anticonvulsant Drugs and Seizure Induction

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PTZ (catalog #P6500), PHT (catalog #PHR1139), methylcellulose (VEH; catalog #M0430), valproic acid (VPA; catalog #P4543), carbamazepine (CBZ; catalog #C4024), potassium permanganate (KMnO4; catalog #223468), xylenes (catalog #XX0060), xylazine (X1251–1G), and DPX mounting medium (catalog #06522) were all from Sigma Aldrich (St. Louis, MO, USA). Lorazepam (LZP; NDC 0641–6046-01) was from Westward Pharmaceuticals as a solution, which was further diluted in 40% hydroxypropyl-β--cyclodextrin. Formal Fixx (catalog #9990244), Hoeschst 33342 (catalog #62249), and NaOH (catalog #SS254) were from Thermo Fisher Scientific (Waltham, MA, USA). Fluoro-Jade C (FJ-C; catalog #1FJC) was from Histo-Chem Inc (Jefferson, AR, USA). Kainic acid (KA; catalog #0222) was from Tocris (Bristol, UK). Ketamine (Ketaset) was from Zoetis, Inc (Parsippany, NJ). Anticonvulsant drugs (PHT; CBZ; VPA) administered to mice were suspended in VEH; PTZ was administered after dissolving in saline.
Knox K.M., Zierath D.K., White H.S, & Barker-Haliski M. (2021). Continuous seizure emergency evoked in mice with pharmacological, electrographic, and pathological features distinct from status epilepticus. Epilepsia, 62(12), 3076-3090.
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8

Cardiac Cell Culture and Imaging

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LPI sodium salt from Glycine max (soybean), CID‐16020046, TTC, PBS, BSA, Cor.At® CL‐i cardiomyocytes (miPSC cardiomyocytes) 1M kit with puromycin and Cor.At® medium, HBSS and all the components for Kreb's Henseleit solution (with the exception of NaCl and NaHCO3) were all purchased from Sigma Aldrich®, UK. Ketamine (Vetalar™) and Xylazine (Rompun®) were obtained from Pfizer and Bayer Healthcare (both Dublin, Ireland) respectively. Y‐27632 dihydrochloride was purchased from Tocris Bioscience (Abingdon, UK). NaCl, NaHCO3, and DMSO were bought from Fisher Scientific, UK. Formal Fixx™ was supplied by Thermo Fisher Scientific, UK. HEPES was bought from Invitrogen, UK. Corning® Epic® 384 Well Cell‐Based Assay Microplates were acquired from Corning Life Sciences, UK. Human iPSC Cardiomyocytes (hiPSC cardiomyocytes; iCell® Cardiomyocytes) were acquired from Cellular Dynamics®, US.
Robertson‐Gray O.J., Walsh S.K., Ryberg E., Jönsson‐Rylander A., Lipina C, & Wainwright C.L. (2019). l‐α‐Lysophosphatidylinositol (LPI) aggravates myocardial ischemia/reperfusion injury via a GPR55/ROCK‐dependent pathway. Pharmacology Research & Perspectives, 7(3), e00487.
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9

Quantifying Calcium and Phosphorous in Scaffolds

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Inductively coupled plasma (ICP) mass spectroscopy was used to assess the amount of calcium and phosphorous produced by hMSCs seeded on scaffold variants (Figure 3C). Briefly, scaffolds were added to Formal-Fixx (10% neutral buffered formalin, ThermoFisher Scientific) for 24 h at 4°C, washed in PBS three times for 5 min each, partially dried via blotting (Kimwipe), stored at -80°C, then dried via lyophilization prior to analysis. ICP optical emission spectrometry (OES) was performed on fixed, dried scaffolds (n = 6 per group). Samples were weighed prior to being dissolved in concentrated nitric acid (Trace Metal Grade concentrated HNO3, Thermo Fischer Scientific 67%–70%) then subjected to automated sequential microwave digestion (CEM Mars 6 microwave digester). The resulting acidic solution was diluted to a volume of 50 ml using DI water to a final concentration <5% acid. ICP-OES was calibrated with a series of matrix matched standards before introducing unknown collagen samples. Digestion and ICP-OES analysis parameters are listed in Supplementary Table S4.
Kolliopoulos V., Dewey M.J., Polanek M., Xu H, & Harley B.A. (2022). Amnion and chorion matrix maintain hMSC osteogenic response and enhance immunomodulatory and angiogenic potential in a mineralized collagen scaffold. Frontiers in Bioengineering and Biotechnology, 10, 1034701.
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10

Characterizing Atherosclerotic Plaque Composition

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Hearts were formalin fixed (3.7% neutral-buffer formalin; Shandon Formal-Fixx, Thermo scientific, Runcorn, UK) and embedded in OCT compound (Optimum Cutting Temperature; Sakura Finetek Europe B⋅V., Alphen aan de Rijn, The Netherlands) before sectioning. Cryosections of 10 μm were collected in Menzel Gläser SuperFrost® Plus slides (Thermo Scientific; USA). Plaque size per valve was determined after staining of cryosections for neutral lipids using Oil Red O and hematoxylin (Sigma-Aldrich, Zwijndrecht, The Netherlands). For the macrophage staining, a primary monoclonal rat-anti-mouse Cd68 antibody (FA-11; ab53444; Abcam, Cambridge, UK) was used after 1:1000 dilution in blocking buffer. A secondary AP-conjugated goat-anti-rat IgG (A8438, Sigma-Aldrich, Zwijndrecht, The Netherlands) was used at a dilution of 1:100 in blocking buffer. The ready-to use BCIP®/NBT liquid substrate system (sigma-Aldrich, Zwijndrecht, The Netherlands) was used for detection. Corresponding sections were stained using the Masson's Trichrome method (Sigma-Aldrich) to determine collagen content. Mean lesion, macrophage, and collagen area (μm [2] ) were quantified using pictures generated with a digital slide scanner (PANNORAMIC 250 Flash II, 3dHistech) and image J software. Quantification was performed blinded.
Zhang Y., Verwilligen R.A.F., de Boer M., Sijsenaar T.J.P., Van Eck M, & Hoekstra M. (2021). PRMT4 inhibitor TP-064 impacts both inflammatory and metabolic processes without changing the susceptibility for early atherosclerotic lesions in male apolipoprotein E knockout mice. Atherosclerosis, 338.
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