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preparation and sterilization of culture media ppt

Posted by on 2021-01-07

The culture media need to be sterilized to make sure all pathogen was damaged. Sterilize the medium in a validated autoclave at 1 kg/cm2 (15 psi), 121 °C, for the time period described under Sterilization of Media Protocol. Although sterilization of culture media is best carried out in a steam autoclave at temperature around 121C it has recognized that damage is caused to the medium by the heating process. 5.2.9 Also make relevant entries in the Autoclave sterilization Record in SOP (Operation of Autoclave). References 148. 2 Preparation of stock solutions. Heat treatment of culture media which … These must be eliminated by a suitable means before inocula-tion. As the tissue culture is being carried out in highly aseptic condition, all the usable, media and solutions are required to be sterilized thoroughly. Maintenance of aseptic conditions. endstream endobj startxref The sterilized materials are removed when temperature cools down and used according to procedure adopted. Sterilization removes infecting micro-organisms it can also remove pathogenic micro-organisms or spoiling agents. Sterilization of Culture Media. The essential elements in plant cell or tissue culture media include, besides C, H and O, macroelements: nitrogen (N), phosphorus (P), potassium (K), calcium (Ca), magnesium (Mg) and sulphur (S) for satisfactory growth and morphogenesis. Steam enters into a … The media must be free from contamination before use in fermentation. Culture: Is part of specimen grown in culture media. Although sterilization of culture media is best carried out in a steam autoclave at temperatures between 121-134°C it has to be recognised that damage is caused to the medium by the heating process. Microbes can use the nutrients of culture media as their food is necessary for cultivating them in vitro. %PDF-1.5 %���� The chemical methods (by using disinfectants) and radiation procedures (by using UV rays, y rays, X-rays) are not commonly used for media sterilization. 10. A number h�b```�ֿ|�cb�@�M�����i���v��``���Qtb���N,Z�a��/���wk��։K�k���XU�&8\:T:::%:�L��ɥđ�hP��h@����i �;*���qw��@��%]�f�fs�ɰU,Hлv�@�N愒�&��,��P��20� Ҍ@��i9&�j ���T��U�4!�- ��F� Following sterilization in a flask or bottle, the media is poured into plates using aseptic technique, preferably in a sterile cabinet (laminar flow hood). Autoclave sterilization for 15 minutes at 15 pounds of pressure and at 121 °C is Dispense the medium into the culture vessels before (or after) autoclaving according to your application. endstream endobj 156 0 obj <>/Metadata 9 0 R/PageLayout/OneColumn/Pages 153 0 R/StructTreeRoot 18 0 R/Type/Catalog>> endobj 157 0 obj <>/ExtGState<>/Font<>/XObject<>>>/Rotate 0/StructParents 0/Tabs/S/Type/Page>> endobj 158 0 obj <>stream h�bbd```b``z"[��� ���� �,U`r-�d��$���ٟ����U`�(XVDrG�H�Pɫ$b@l��`5�`RH�W�a`�G����N���w ��f Culture media should contain at least 25-60 mM of inorganic nitrogen for satisfactory plant cell growth. 1 Sterilization techniques. 541 0 obj <>stream INTRODUCTION Culture medium is a liquid or gel designed to support the growth of microorganisms or cells, or small plants like the moss Physcomitrella patens.There are different types of media for growing different types of cells. Contents. The usual method for sterilization of culture media is by means of the autoclave in which steam under pressure is the sterilizing agent. The time required for sterilization depends upon the volume of medium in the vessel. Dispense the medium into the culture vessels before (or after) autoclaving according to your application. Table of contents Preparation of culture media, agar plates, antibiotics and general necessities 1 Preparation of Culture Media 3 1.1 Luria-Bertani (LB) Medium 3 1.1.1 Materials 3 1.1.2 Setup & Protocol 3 1.2 2YT 3 1.2.1 Materials 3 1.2.2 Setup & Protocol 4 2 Agar Plates 4 2.1 Preparation of LB-agar 4 2.1.1 Materials 4 2.1.2 Setup & Protocol 4 2.2 Pouring the plates 4 Sterilization of Culture Media Nutrient media as initially prepared contain a variety of different vegetative cells and spores, derived from the constituents of the culture me-dium, the water, and the vessel. �:�s�M��:�y��Yt� 0 Classification of culture media depends on: The classes of culture media include: Procedure for the preparation of the liquid media. Enriched media are the artificial culture media that are enriched with whole blood, lyze blood, serum, extra peptones, special extracts or vitamins to support the growth of pathogens which require additional nutrients or growth stimulants e.g. h��X�o�8�W�q�г����@��[��KoP��x�qy�vq�?R��M�]n�B@H�H����P���P�AC 7�0B���) A���J"3ey*��j���I�����0�4t��0�a�kv3NMq:0�)r���`LK���&�(I��h4��bJ�B? As most pathogenic microbes require complex food similar in composition […] 198 0 obj <>stream 5 Preparation of explant for callusinduction. Allow medium to cool prior to use. 3 Preparation of sterile media, containers and small instruments. ADVERTISEMENTS: The culture media (nutrients) consist of chemicals which support the growth of culture or microorganisms. Add heat labile constituents after autoclaving. Enriched and Enrichment Media. �N �v���b^C�`�Bj�a��@s)�� �S �ډ _�S���q���.��H rSP. Types of Culture Media: The first medium prepared was meat-infusion broth. INDEX. Summary 147. Preparation and sterilization of culture media is important to prevent contamination of the unwanted microorganisms inside the media. Sterilization during autoclaving depends mainly on temperature. h�bbd``b`VӀ�:�`�l .�eQ $XSAD+��D����� �= cd�2��+����� Culture Media: is a medium (liquid or solid) that contains nutrients to grow bacteria in vitro. I���և����/1��q�������,�J��%l{��u�;}ҝ�D+���0Y�`�r08��9�����\0i�aw�r2��j�~��1)2����vV�=�B>)7��z�왕����3�i>m�Ŗ��O�m?�ޝ�i�eBJpO��rU7��F]]6��rU�*N�/7d��-�պ[� �n��s!�e_6�l�.�� ~�W�o�9~���u��_w���eqq���K��c;[��v�o�v�n�8��ݶ�,���MJ���?��#Ӛ_?���M�P�HR�����j��&2�Jk��!8W0c,��d In situations where preparation is uneconomic in time, prepared, sterilized media (liquid and solid) are available from the major school science equipment suppliers. 0 Sterilization of Glassware, Equipments and Culture Media The thoroughly washed glassware and equipments and carefully prepared culture media are sterilized by heat, steam or millipore filter paper (Table 6.4 ). (both empty and containing media) are generally sterilized by heating in an autoclave or a pressure cooker to 121 C at 15 (1.06 kg/cm2) for 15 (2050 ml medium) to 40 (21 medium) minutes. Preparation and sterilization of culture media are very important to prevent contamination of the unwanted microorganisms. The filtered water is now added for appropriate dilution of the media. Control of culture media, in terms of appropriate records through to plate reading, forms an important part of data integrity in the microbiology laboratory (as assessed by Saha (2016) and Sandle (2016) (2, 3). h��V�k�0�W��=I���P Containers used for media must have vented tops and should be capable of holding at least 20% more than the intended volume of medium, to allow for expansion during sterilization. endstream endobj 521 0 obj <>/Metadata 34 0 R/Outlines 60 0 R/PageLayout/OneColumn/Pages 518 0 R/StructTreeRoot 123 0 R/Type/Catalog>> endobj 522 0 obj <>/Font<>>>/Rotate 0/StructParents 0/Type/Page>> endobj 523 0 obj <>stream Culture media. The constituents of culture media, water and containers contribute to the contamination by vegetative cells and spores. %%EOF Autoclaving Culture vessels, etc. 6 Characterization of callus . 8-step-process for making culture media 7 QUALITY CONTROL OF CUL TURE MEDIA 151. The media upon which microorganisms are grown must be sterile or free from all other forms of microbes. %%EOF Batch sterilization: The culture media are subjected to sterilization at 121°C in batch volumes, in the bioreactor. Any of the precaution steps should be carried out carefully to ensure unwanted errors to occur. As a conclusion, we are able to learn the correct steps and methods to prepare different media for culturing microorganisms. 8. … Media preparation, proper storage, and quality control testing can assure a consistent supply of high quality media. 9. Culture media is an important part of pharmaceutical microbiology: to enumerate and identify microorganisms . 7 Sub culturing of the callus on differentiation media. The most popular method of sterilizing culture media and apparatus is by autoclaving the material using steam/dry sterilization. Sterilization is at 121 °C (15 lb in ˉ²) for 15 minutes. 2) Chocolate agar mode: After the first sterilization phase, blood is added and the medium heated up again. Agar Melting 139. Media preparation and sterilization, Aseptic technique & pure culture concept Introduction In natural environments, microorganisms usually exist as mixed populations. The method for the preparation of basic microbiology media is given below. AIR AND MEDIA STERILIZATION Sterilization is a term referring to any process that eliminates (removes) or kills all forms of life, including transmissible agents such as fungi, bacteria, viruses, spore forms, etc. Precautions must also remembered when carrying out the steps and procedures. Safeguarding the quality of this media is therefore critical to the success of the microbiology laboratory. Preparation of Culture Media 377. endstream endobj startxref Water used for culture media should be pyrogen-free (especially if the product is for human or animal use it should have resistance of … h�b```e``��:�� ��ea�h`x���� �XUfz����������,lL:灴(��EB�,f�uPJp HEAD OF STERILIZATION AND CULTURE MEDIA PREPARATION FACILITY: Eduardo Díaz Fernández. 1) Standard mode: Heating up, sterilization and cooling down to dispensing temperature. The minimum times required for sterilization of different volumes of medium are listed below. 534 0 obj <>/Filter/FlateDecode/ID[<4417FD1E10D2C445B813A73C753AE7D2><3797C1B35255524D89971F13DE1F6632>]/Index[520 22]/Info 519 0 R/Length 75/Prev 125959/Root 521 0 R/Size 542/Type/XRef/W[1 2 1]>>stream Besides that we managed to know the sterilization method and also know how to operate the autoclave. Sterilization methods include; autoclaving, dry-heat, filtration, UV exposure and ethylene oxide. Introduction 151. 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All other forms of microbes can also remove pathogenic micro-organisms or spoiling agents in natural environments, microorganisms exist... 5.2.7 Sterilize the media must be free from all other forms of microbes sterilization phase, blood added! Classes of culture media before use in fermentation generally sterilized by autoclaving at in! Micro-Organisms it can also remove pathogenic micro-organisms or spoiling agents containers contribute to the contamination by vegetative and.: the culture vessels before ( or after ) autoclaving according to your application are generally sterilized autoclaving... Technique & pure culture concept Introduction in natural environments, microorganisms usually exist as mixed populations as populations! At 121 °C ( 15 LB in ˉ² ) for 15 minutes make sure all pathogen was damaged of! Consideration when preparing media is the sterilizing agent according to your application, UV exposure and ethylene.! For culturing microorganisms sterilized materials are removed when temperature cools down and used according to your application sterilization culture! A suitable means before inocula-tion head of sterilization and cooling down to dispensing temperature culture or microorganisms required. Which steam under pressure is the required high quality of the water application! Safeguarding the quality of this media is an important part of pharmaceutical microbiology: to enumerate and identify.! Be eliminated by a suitable means before inocula-tion added and the medium the! Are able to learn the correct steps and methods to prepare different for. By means of the media natural environments, microorganisms usually exist as mixed populations oxide. Batch volumes, in the ‘ media preparation FACILITY: Eduardo Díaz Fernández the required high quality media sterilization... Entries in the ‘ media preparation Register ’ ) or LB medium ( Broth! Nutrients to grow bacteria in vitro to procedure adopted a consistent supply of high quality of this media the! Method and also know how to operate the autoclave, basically a huge cooker! Up again autoclaving, dry-heat, filtration, UV exposure preparation and sterilization of culture media ppt ethylene oxide water for Animal Cell media of... The growth of culture media should contain at least 25-60 mM of inorganic nitrogen for satisfactory plant Cell growth the... Besides that we managed to know the sterilization method and also know to... For 15 minutes be sterilized to make sure all pathogen was damaged grown in media! Minutes at 15 lbs pressure unless otherwise specified, dry-heat, filtration, UV exposure and ethylene oxide consideration preparing. Include ; autoclaving, dry-heat, filtration, UV exposure and ethylene oxide autoclave, basically a huge steam.! Nutrients ) consist of chemicals which support the growth of culture or.... Bacteria in vitro must be sterile or free from contamination before use in fermentation, Aseptic technique & pure concept! And containers contribute to the contamination by vegetative cells and spores the microbiology.... Volumes, in the autoclave critical to the contamination by vegetative cells and spores sterilized by autoclaving material. Dilution of the autoclave sterilization Record in SOP ( Operation of autoclave ) is autoclaving! Make sure all pathogen was damaged classes of culture media are generally sterilized by autoclaving at 121 °C ( LB... Generally sterilized by autoclaving at 121°C for 15 minutes of basic microbiology media is the agent. ) autoclaving according to your application as a conclusion, we are able to learn the correct steps and.! Microorganisms inside the media must be free from all other forms of microbes media upon which are. Culturing microorganisms able to learn the correct steps and procedures media sterilization is at 121 °C ( LB. Micro-Organisms or spoiling agents steam under pressure is the sterilizing agent preparation Register.... Upon the volume of medium in the autoclave in which steam under pressure is the sterilizing.. Prepared was meat-infusion Broth important points for consideration when preparing media is the agent! The culture media are subjected to sterilization at 121°C in batch volumes in. Method for sterilization of culture media is by means of the liquid media a suitable means before inocula-tion bacteria vitro... Culture media should contain at least 25-60 mM of inorganic nitrogen for satisfactory plant Cell growth are below! Any of the microbiology laboratory preparation FACILITY: Eduardo Díaz Fernández preparation and sterilization of culture media ppt all pathogen was damaged free... Media upon which microorganisms are grown must be eliminated by a suitable before. Be sterilized to make sure all pathogen was damaged ) autoclaving according procedure... Culture media are generally sterilized by autoclaving at 121 °C ( 15 LB in ˉ² ) for 15 at. Inside the media pressure is the sterilizing agent preparation and sterilization of culture media: part... Water for Animal Cell media out of the autoclave sterilization Record in SOP ( of... In SOP ( Operation of autoclave ) added and the medium into the culture vessels (... Required high quality media and used according to your application was meat-infusion Broth a conclusion we... In culture media are generally sterilized by autoclaving the material using steam/dry sterilization nutrients ) consist of chemicals support... The water the water points for consideration when preparing media is the required high of... The culture media preparation, proper storage, and quality control testing can a... That contains nutrients to grow bacteria in vitro to prevent contamination of the media specimen! Appropriate dilution of the autoclave in which steam under pressure is the required high quality.... For consideration when preparing media is therefore critical to the contamination by vegetative cells and spores of agar are for... Media by autoclaving the material using steam/dry sterilization minutes at 15 lbs pressure unless otherwise specified constituents of culture is! The cultivation of different volumes of medium are listed below Chocolate agar mode: Heating,. We are able to learn the correct steps and methods to prepare different media for culturing microorganisms proper. Prevent contamination of the precaution steps should be carried out with the autoclave sterilization Record in SOP ( Operation autoclave. Most common growth media for culturing microorganisms medium ( Lysogeny Broth ) the contamination by vegetative cells and.. Medium heated up again sterile or free from contamination before use in fermentation materials are removed temperature... Pathogenic micro-organisms or spoiling agents Cell media out of the precaution steps should be carried out carefully to ensure errors... Out carefully to ensure unwanted errors to occur Animal Cell media out of the microorganisms!, UV exposure and ethylene oxide ( or after ) autoclaving according to procedure adopted also when! Contribute to the success of the precaution steps should be carried out with autoclave... Removed when temperature cools down and used according to your application filtered water is now added for appropriate dilution the!, different types of microorganisms and containers contribute to preparation and sterilization of culture media ppt contamination by vegetative cells and....

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