cryotechniques by priya garg holkar science college indore

Govt. [Model, Autonomous] Holkar Science College, Indore

शा. (आदश�, स्वशासी) होलकर विवज्ञान महाविवद ्यालय, इन्दौर

2015-16Department of Zoology

Guided byDr. ABIDA QURESHIMADAM

Submitted byPRIYA GARG DS1290945M.Sc. 2 Semester

SYNOPSIS INTRODUCTION TYPES CRYOPRESERVATION OF CELLS FREEZABLE TISSUES CRYOPRESERVATION OF EMBRYO CRYOSURGERY USES OF CRYOSURGERY INSTRUMENTS OF CRYOSURGERY CRYOTOMY FREEZE FRACTURE FREEZE DRYING

CRYOTECHNIQUES• Cryotechniques are a group of related

procedures for stabilizing, or “fixing” specimens for microscopic observation.

• Samples are rapidly frozen to maintain cellular structure and composition as it exists under physiological conditions, such as electrolyte concentration and protein antigenicity.

• Alternatively, samples may be lightly fixed and cryoprotected first before freezing.

• These techniques preserve the native structures of tissues without the artifacts associated with chemical fixation.

• Some cryotechniques facilitate investigations of membranes and membrane proteins.

CRYOTECHNIQUES

CRYOPRESERVATION OF CELLS, TISSUES, ORGANS AND ORGANISMS

CRYOSURGERY

CRYOTOMY

FREEZE FRACTURE

FREEZE DRYING

CRYOPRESERVATION OF CELLS, TISSUES, ORGANS AND ORGANISMS

•Cryopreservation or cryoconservation is a process where cells,  tissues, or any other substances susceptible to damage caused by chemical reactivity or time are preserved by cooling to sub- zero temperatures.

• At low enough temperatures, any enzymatic or chemical activity which might cause damage to the material in question is effectively stopped.

•Cryopreservation methods seek to reach low temperatures without causing additional damage caused by the formation of ice during

freezing.

Freezable tissuesLivers hearts :Semen in semen cryopreservationBloodStem cells Umbilical cord blood Tissue samples like tumors and histological cross sectionsEggs (oocytes) in oocyte cryopreservationEmbryos at cleavage stage (that are 2, 4 or 8 cells) or at        

blastocyst stage, in embryo cryopreservationOvarian tissue in ovarian tissue cryopreservationPlant seeds or shoots may be cryopreserved  for 

conservation purposes.

Laboratory for cryopreservation

Nitrogen tub

Cryopreserved brain

Human body being cryopreserved

Dead body - cryopreserved

Cryopreservation of semen

CryosurgeryCryosurgery (cryotherapy) is the use of extreme cold in surgery to destroy abnormal or diseased tissue.

The term comes from the Greek words cryo   ("icy cold") and surgery meaning "hand work" or "handiwork".

Cryosurgery has been historically used to treat a number of diseases and disorders, especially a variety of benign and malignant skin conditions.

UsesWarts, moles, skin tags, solar keratoses, Morton's neuroma[3] and

small skin cancers are candidates for cryosurgical treatment. Several internal disorders are also treated with cryosurgery, including liver cancer, prostate cancer, lung cancer, oral cancers, cervical disorders and, more commonly in the past, hemorrhoids. Soft tissue conditions such asplantar fasciitis (Jogger's heel) and fibroma(benign excrescence of connective tissue) can be treated with cryosurgery. Generally, all tumors that can be reached by the cryoprobes used

during an operation are treatable.Although found to be effective, this method of treatment is only appropriate for use against localized disease, and solid tumors larger than 1 cm. Tiny, diffuse metastases that often coincide with cancers are usually

not affected by cryotherapy.

Method Liquid nitrogen Cryogun used to spray liquid nitrogen

Carbon dioxideCarbon dioxide is also available as a spray and is used to treat a variety of benign spots. Less frequently, doctors use carbon dioxide"snow" formed into a cylinder or mixed with acetone to form a slush that is applied directly to the treated tissue.

ArgonRecent advances in technology have allowed for the use of argon gas to drive ice formation using a principle known as the Joule-Thomson effect.

Dimethyl ether – propaneA mixture of dimethyl ether and propane is used in some preparations such as Dr. Scholl's Freeze Away.

Some commonly used cryosurgical products are:BrymillCry-AcCryoalfaCryoClearCryoPenCryoPro, Cortex TechnologyCryoProbeCryosurgery, Inc. Verruca-FreezeHistofreezerMedGyn Cryotherapy SystemMiltex CryosolutionsPremier CryOmegaPremier NitroSprayMyoscience Iovera

ResultsCryosurgery is a minimally invasive procedure, and is often preferred

to more traditional kinds of surgery because of its minimal pain, scarring, and cost; however, as with any medical treatment, there are risks involved, primarily that of damage to nearby healthy tissue.

Damage to nerve tissue is of particular concern.

Patients undergoing cryosurgery usually experience redness and minor-to-moderate localized pain, which most of the time can be alleviated sufficiently by oral administration of mild

analgesics such as Ibuprofen, codeine, tramadol or acetaminophen (paracetamol Blisters may form as a result of cryosurgery, but these usually scab

over and peel away within a few days.

cryosurgery · benign keratosis · benign keratosis

Cryosurgery: Frozen Hazard for Cancer | Healthy Panacea

Skin Cancer Treatment – Cryosurgery: A treatment that uses an instrument to freeze and destroy abnormal tissue, such as carcinoma in situ. This type of treatment is also called ...

CryotomyTissue cut into thin sections.Frozen sections are a method of tissue analysis that allows for rapid interpretation and diagnosis of tissue during surgery. Cryotomy can also be used in the

preparation of sections containing fats and enzymes which can easily be lost in alcohol or paraffin

sections.

Freeze FractureIn the freeze fracturing process, a specimen is frozen rapidly and cracked on a plane through the tissue. This fracture occurs along weak portions of the tissue such as membranes or surfaces of organelles.After cleaving, both surfaces are shadowed with a platinum film. This coating produces a replica of the surfaces. The replica is then coated with carbon and is then imaged in the transmission electron microscope.                                                      The method is also good for the study of intramembrane structures. 

Freeze-drying—is a dehydration process typically used to preserve a perishable material or make the material more convenient for transport. Freeze-drying works by freezing the material and then reducing the surrounding pressure to allow the frozen water in the material to sublimate directly from the solid phase to the gas phase.

The freeze-drying stagesPretreatmentFreezingPrimary dryingSecondary dryin

g

Primary dryingDuring the primary drying phase, the pressure is lowered and enough heat is supplied to the material for the ice to sublime. The amount of heat necessary can be calculated using the sublimating molecules’ latent heat of sublimation.

Secondary dryingThe secondary drying phase aims to remove unfrozen water molecules, since the ice was removed in the primary drying phase. This part of the freeze-drying process is governed by the material’s adsorption isotherms.

Applications of freeze-dryingPharmaceutical companies often use freeze-drying to increase the shelf life of the products, such as vaccines and other injectables. By removing the water from the material and sealing the material in a vial, the material can be easily stored, shipped,and later reconstituted to its original form for

injection.

Image of a freeze dryer

Image of a freeze dryer

Image of a freeze dryer

REFERENCESMICROBIOLOGY BY MICHAEL . J. ` PELCZARBIOTECHNOLOGY BY B.D. SINGHTECHNIQUES IN MICROSCOPY

AND CELL BIOLOGY BY V.K . SHARMA

Wikipedia, the free encyclopedia

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