Uranyless

Uranyless Aqueous

Uranyless aqueous is a contrasting solution that substitutes for uranyl acetate. As recommended by Reynolds (1963), it is highly advisable to reinforce the initial contrast (in our case, UranyLess) with lead citrate in a NaOH-saturated atmosphere to prevent contamination from atmospheric CO2.

To facilitate the use of lead citrate during contrasting, it is packaged in AirLess bottles (without air and thus without CO2). You can store this 30 ml bottle for a long time without the risk of CO2 contamination, a common problem for microscopists.

The 30 ml AirLess bottle packaging allows you to contrast over 1500 grids.

This packaging is doubly effective: it provides long shelf life and generates minimal waste.

For using UranyLess aqueous in automated contrast devices like Leica’s EM Stain, UranyLess is available in a 200 ml bottle. It is worth noting that UranyLess aqueous is packaged in an AirLess bottle to facilitate drop dispensing only, as it is not affected by air or light.

Yeast

Preparation of the sample according to the following protocol:

Standard fixation with Glutaraldehyde – Osmium – Embedding in Epon.
Contrast with UranyLess followed by Lead Citrate.

Here are some results from Jeannine Lherminier (INRA – Dijon).

Photographs taken using Transmission Electron Microscopy

Trematodes

We present some results from Yann Quilichini (Microscopy Platform at the University of Corsica – Corte).

Sample preparation according to the following protocol:

Standard fixation with glutaraldehyde, osmium, embedding in Spurr resin.
Thin sections – contrast with aqueous UranyLess followed by lead citrate (according to Reynolds).

Photographs taken using Transmission Electron Microscopy.

Polymersomes-Uranyless

The IMRCP Laboratory in Toulouse, led by Anne-Françoise Mingotaud, tested UranyLess in comparison to uranyl acetate, which is at an acidic pH of 4 (seems to disrupt the molecular structure organization). They also compared observations using Cryo-SEM (scanning electron microscopy).

The chemical structure is organized as follows:

Observation using Scanning Electron Microscopy in Cryo mode.

Reconstructed Epidermis

We present the results from Audrey Houcine (CMEAB Toulouse).

Sample preparation according to the following protocol:

Standard fixation with glutaraldehyde, osmium, Epon/Araldite
Ultrathin sectioning, double contrast with Uranyless followed by lead citrate

Photographs taken using Hitachi HT7700 Transmission Electron Microscopy by Audrey Houcine.

Muscle-Nerve-Mince

Sample preparation according to the following protocol:

Standard fixation with glutaraldehyde, osmium, Epon
Ultrathin sectioning, double contrast with Uranyless 1min followed by lead citrate 1min

Mouse Ovarian Follicle

Sample preparation according to the following protocol:

Standard fixation with glutaraldehyde Fixative PFA, osmium, Epon
Ultrathin sectioning, double contrast with Uranyless 1min followed by lead citrate 1min

Mouse Kidney

Mouse ovarian follicle

Sample preparation according to the following protocol:

Standard fixation with glutaraldehyde Fixative PFA, osmium, Epon
Ultrathin sectioning, double contrast with Uranyless 1min followed by lead citrate 1min

Kidney

Mouse Kidney

Sample preparation according to the following protocol:

Classic PFA- Glutaraldehyde, Osmium, Epon fixing.
Ultrafine section – 1 minute UranyLess contrast – 1 minute Lead Citrate.

Photos taken with a Hitachi HT7700 Transmission Electron Microscope by Nacer BENMERADI (R&D – Delta Microscopies – France).

Cardiac muscle - Uranyless

Mouse cardiac muscle

Preparation of the sample using following protocol:

classic Fixing Glutaraldehyde, Osmium, Epon Ultrafine
Cups, Double Uranyless Contrast and Lead Citrate

Plant Tissu

Mouse cardiac muscle

Preparation of the sample using following protocol:

classic Fixing Glutaraldehyde, Osmium, inclued in epon
Double Uranyless Contrast and Lead Citrate

Phage T6

Sample preparation according to the following protocol:

Staggering Phage t6 on a G300-Cu grid covered with formvar film. Lonization 1 min.

Cross sectional Bacteria/Bacteri E-Coli

Sample preparation according to the following protocol: Cross sectional Bacteria

Fixing Classic Glutaraldehyde Osmium, EPON
Cutting ultrafine, double contarst Uranyless and lead Citrat

Bacteri E-coli

Negative Staining for 2 minutes Uranyless bacteria like E-coli (adherent and invasive (ACSI) LF82) Which have Pili and Flagella .

Sacculina crustacean/Intestine

Sample preparation according to the following protocol: Sacculina Crustace (small parasitic crustacean)

Fixing Classic Glutaraldehyde Osmium, EPON inclusion
Fines Cups- Contrast to the aqueous Uranyless 60°C on a Hotplate without Lead Citrat post coloring