X 34 dye

Manufactured by Merck Group

The X-34 dye is a laboratory reagent used for staining and visualization purposes. It is a synthetic dye that exhibits fluorescent properties when exposed to specific wavelengths of light. The X-34 dye can be utilized to label and detect various biological molecules, cellular structures, or other targets during scientific research and analysis.

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Lab products found in correlation

Dmd108 microscope, by Leica camera (1 mentions) Dm2500 fluorescent microscope, by Leica camera (1 mentions) Oil red o, by Merck Group (1 mentions) Axioscope fluorescent microscope, by Zeiss (1 mentions)

2 protocols using x 34 dye

Staining Amyloid and Lipid Deposits in C. elegans

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X‐34 dye (Sigma‐Aldrich‐Merck) stains specifically amyloid deposits. One‐day synchronised adults were incubated in a drop containing 1 mM X‐34 and 10 mM TRIS pH 7.5 solution. Stained animals were washed with PBS‐Tween and transferred to fresh NGM plates. We used the Oil Red O (Sigma‐Aldrich‐Merck) dye to stain triglycerides and lipoproteins for semi‐quantitative purposes. Young adults were stained with diluted 3:2 Oil Red O stock solution for 2 h at room temperature in a rotator. After this, animals were washed two times with PBS‐Tween (0.1%) for 30 min. Stained animals were mounted in 2% agarose pads with sodium azide (0.05 M) to count amyloid deposits using a DM2500 fluorescent microscope (Leica). To quantify the stained area of worms, with the Oild Red O stain, we used a DMD108 microscope (Leica). We analysed the pictures obtained from these animals using the Image J software.

Gómez‐Escribano A.P., Mora‐Martínez C., Roca M., Walker D.S., Panadero J., Sequedo M.D., Saini R., Knölker H., Blanca J., Burguera J., Lahoz A., Cañizares J., Millán J.M., Burton N.O., Schafer W.R, & Vázquez‐Manrique R.P. (2023). Changes in lipid metabolism driven by steroid signalling modulate proteostasis in C. elegans. EMBO Reports, 24(6), e55556.

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Live staining and quantification of protein aggregates

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Staining was performed following the protocol by Link et al. ^{100 (link), 101 (link)}. A subset of worms from ongoing paralysis assays was removed from the experiment 24 hours after temperature upshift for live staining. The live nematodes were transferred with OP50 onto 10 μL drops of 1 mM X-34 dye (Sigma-Aldrich) in 10 mM Tris pH 8.0 and stained for 3 hours at 25 °C in a humidity chamber. After staining, animals were transferred to seeded NGM plates and allowed to recover for 4 hours at 25 °C. Worms were then mounted onto 2 % agarose pads with 0.2 % levamisole on glass microscope slides for imaging. Images were captured using a Zeiss Axioscope fluorescent microscope, 400X magnification, BP 365/12 filter set. Aggregates were quantified by counting the number of stained aggregates in the head region, anterior to the pharyngeal bulb ^{102 (link), 103 (link)}. Aggregates vary in size, only overall quantity of aggregates per worm was scored.

Doens D., Valdés-Tresanco M.E., Vasquez V., Carreira M.B., De La Guardia Y., Stephens D.E., Nguyen V.D., Nguyen V.T., Gu J., Hegde M., Larionov O.V., Valiente P.A., Lleonart R, & Fernández P.L. (2019). Hexahydropyrrolo[2,3-b]indole compounds as potential therapeutics for Alzheimer Disease. ACS chemical neuroscience, 10(10), 4250-4263.

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