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Cryogenic sample preparation laboratory (LCRIO)

The Cryogenic Sample Preparation Laboratory (LCRIO) is part of the Biological Sample Preparation Village* and aims to support the preparation of soft and biological materials at room temperature and at low temperatures (-20°C to -190°C), to preserve their native structure and composition during Sirius’ beamlines experiments. The LCRIO is located on axis 40, along the Cateretê beamline.

The LCRIO has equipment that allows users to section, extract and vitrify samples on a micrometric scale, at room temperature, and at low temperatures.

*The Biological Sample Preparation Village aims to support internal and external users in the preparation of biological samples, as well as in conducting sectioning, syntheses, and characterizations for biological studies.

CONTACT & STAFF

Contact: Carla Cristina Polo
Tel.: +55 19 3512 5193
E-mail: carla.polo@lnls.br

MICRO-CUTTING AND MICROMANIPULATION

LCRIO has sample cutting/sectioning and manipulation tools for nano-tomography and nano-spectroscopy experiments.

Semiautomatic rotary microtome (Leica HistoCore Autocut)

Allows sectioning at room temperature, ranging from 0.5 and 500 µm in thickness, from samples embedded in solid media such as paraffins and resins.

It has support for stainless steel and glass knives.

Access specifications here.

Laser microdissection microscope (Leica LMD7)

Allows to cut and extract micro-fragments including soft materials (i.e. cells, plant tissues, polymers) to harder materials (i.e. thin metallic plates).

In addition, it is an optical microscope with brightfield, polarized light, DIC and fluorescence capabilities.

Access specifications here.

Stereo microscope (Olympus SZ61)

Having a long working distance, it allows the visualization and, at the same time, the manipulation of microsamples, either for manual cutting or for transferring to different substrates.

PREPARATIONS AT LOW TEMPERATURES

Radiation damage is an intrinsic phenomenon resulting from the interaction between soft matter and X-rays, and sample preservation at low temperatures (below -20°C) is mandatory to collect reliable and high-resolution data.

Manual cryostat (Leica CM1860 UV)

Allows sectioning at temperatures down to -30°C, between 0.5 and 500 µm thick of samples embedded in cryoprotectant solution (i.e. OCT, glycerol, glucose, etc.)

Access specifications here.

Plunge-freezer (EMS-002)

Allows rapid freezing of microsamples on specific substrates used at the beamlines. Operates with propane/ethane at liquid nitrogen temperatures (-190°C) and allows to achieve freezing in an amorphous ice state of samples up to ~15 µm thick, avoiding the formation of crystalline ice.

Access specifications here.

More

In addition, the LCRIO has a fume hood for chemical manipulations, a refrigerator with a freezer (4°C and -20°C), and a nitrogen reservoir (Dewar) (-190°C) for the temporary storage of samples.

GENERAL RULES FOR USING THE LCRIO INFRASTRUCTURE

  • On-site training and use of PPE (Personal Protective Equipment).
  • To access the LCRIO (Cryogenic Sample Preparation Laboratory), it is mandatory to wear lab coat (long sleeve), safety glasses and nitrile gloves. In addition to PPE, it is compulsory to wear suitable clothing (long pants and fully closed shoes that cover the back of the foot).
  • Declare all samples and reagents on the Safety Form according to the submitted proposal.
  • Chemical reagents and samples not declared on the Safety Form, as per the submitted proposal, will be prohibited. Changes and additions must be formalized in advance and authorized by the Safety Assessment Committee.
  • Only bring to the experimental facilities (beamlines) the materials necessary to conduct the experiments.
  • Reagents and samples should be temporarily stored for the duration of the experiments. After this period, the materials must be removed by the respective user.
  • Materials without proper identification will not be accepted. All samples and chemical reagents must be properly identified. Identification must be legible. Toxic or volatile materials will be stored in the fume hood.
  • Keep the LCRIO in working order, ensuring its organization and internal operating rules in accordance with safety guidelines and recommendations.
  • Only trained users, in person, will be authorized and have access to the LCRIO.