{ "experiment": { "id": 440, "name": "Microbiological sampling_QA", "description": "This template will guide you on how to do monitor microbiological contamination with contact plates, settle plates and swabs. Quality assurance (QA) is a process, where primarily concerns are controls of errors in the performance of tests and verification of test results. All materials, equipment and procedures must be adequately controlled. \r\nThe objectives of microbiological sampling are to allow statements of density, types and locations of microorganism which reside on different surfaces.", "project_id": 223, "created_by_id": 202, "last_modified_by_id": 202, "archived": false, "archived_by_id": null, "archived_on": null, "restored_by_id": null, "restored_on": null, "created_at": "2018-11-15T09:56:28.245Z", "updated_at": "2019-02-19T14:12:34.714Z", "uuid": "c35c71c3-2f8b-45fb-8064-f14e5d4cc724" }, "my_modules": [ { "my_module": { "id": 2192, "name": "Plate preparation", "due_date": null, "description": "", "x": 0, "y": 16, "my_module_group_id": 1250, "created_at": "2018-11-15T09:56:30.961Z", "updated_at": "2019-02-19T14:12:22.351Z", "archived": false, "archived_on": null, "created_by_id": 202, "last_modified_by_id": 202, "archived_by_id": null, "restored_by_id": null, "restored_on": null, "workflow_order": 0, "experiment_id": 440, "state": "uncompleted", "completed_on": null }, "outputs": [ { "id": 6284, "input_id": 2188, "output_id": 2192 }, { "id": 6285, "input_id": 2190, "output_id": 2192 }, { "id": 6286, "input_id": 2194, "output_id": 2192 } ], "my_module_tags": [], "task_comments": [], "my_module_repository_rows": [], "user_my_modules": [], "protocols": [ { "protocol": { "id": 3548, "name": null, "authors": null, "description": null, "added_by_id": 202, "my_module_id": 2192, "team_id": 1, "protocol_type": "unlinked", "parent_id": null, "parent_updated_at": null, "archived_by_id": null, "archived_on": null, "restored_by_id": null, "restored_on": null, "created_at": "2018-11-15T09:56:31.020Z", "updated_at": "2019-01-31T09:33:37.119Z", "published_on": null, "nr_of_linked_children": 0 }, "protocol_protocol_keywords": [], "steps": [ { "step": { "id": 4533, "name": "Examine the plates", "description": "\n
Follow the guidelines bellow.
", "position": 0, "completed": false, "completed_on": null, "user_id": 202, "created_at": "2018-11-15T09:56:31.091Z", "updated_at": "2018-12-24T08:23:09.402Z", "last_modified_by_id": 202, "protocol_id": 3548 }, "checklists": [ { "checklist": { "id": 924, "name": "Guideline:", "step_id": 4533, "created_at": "2018-12-12T07:16:14.619Z", "updated_at": "2018-12-24T08:23:09.400Z", "created_by_id": null, "last_modified_by_id": null }, "checklist_items": [ { "id": 3812, "text": "Monitor refrigerator temperature and record it in Results.", "checked": false, "checklist_id": 924, "created_at": "2018-12-12T07:16:14.622Z", "updated_at": "2018-12-12T07:16:14.622Z", "created_by_id": null, "last_modified_by_id": null, "position": 0 }, { "id": 3813, "text": "Contact plates and/or Settle plates are taken to the room temperature. They will be used depending on the surface of the sampling.", "checked": false, "checklist_id": 924, "created_at": "2018-12-12T07:16:14.634Z", "updated_at": "2018-12-12T07:16:14.634Z", "created_by_id": null, "last_modified_by_id": null, "position": 1 }, { "id": 3814, "text": "Examine the plates for contamination prior to use. If any plates are contaminated they should be discarded according to protocol.", "checked": false, "checklist_id": 924, "created_at": "2018-12-12T07:16:14.641Z", "updated_at": "2018-12-12T07:16:14.641Z", "created_by_id": null, "last_modified_by_id": null, "position": 2 } ] } ], "step_comments": [], "step_assets": [], "assets": [], "step_tables": [], "tables": [] }, { "step": { "id": 4535, "name": "Preincubation", "description": "\nPreincubate the empty plates in incubator 30°C-35°C for 48 hours to check for any accidental contamination. Check for contamination again. If there is contamination record it and discard according to appropriate procedures. Take contamination-free plates to room temperature an hour before exposure.
All samples (contaminated or not) should be disposed of according to procedures.
", "position": 2, "completed": false, "completed_on": null, "user_id": 202, "created_at": "2018-12-12T11:23:59.242Z", "updated_at": "2018-12-12T11:24:31.752Z", "last_modified_by_id": 202, "protocol_id": 3564 }, "checklists": [], "step_comments": [], "step_assets": [], "assets": [], "step_tables": [], "tables": [] }, { "step": { "id": 5173, "name": "Counting colonies and recording", "description": "\nAfter appropriate incubation microbiological contamination should grow into discrete macroscopic colonies that can be enumerated and the number of discrete colonies forming units (CFU) can be counted on each sample.
Record the number (per unit surface area) on the results tab. Separate colony counts may be tabulated for mould and bacteria.
Colony types may be identified if this is considered appropriate.
Assemble the plates required and label them. Ensure that correct information about sampling is written on the base of the plate with a permanent ink marker.
The following sampling details should be recorded on the plate:
After all the samples are taken, collect all sample swabs together, remove them from the sampling area and plate them as directed in the next step. Complete and enclose all the necessary documentation in [#Counting and recording...~tsk~ZU].
", "position": 2, "completed": false, "completed_on": null, "user_id": 202, "created_at": "2018-11-15T09:56:30.601Z", "updated_at": "2019-01-25T12:40:08.288Z", "last_modified_by_id": 202, "protocol_id": 3544 }, "checklists": [], "step_comments": [], "step_assets": [], "assets": [], "step_tables": [], "tables": [] }, { "step": { "id": 4521, "name": "Sampling with a swab", "description": "\nSampling: Wipe the swab over the sample area in close parallel streaks, using firm even pressure and rotating the swab between fingers to maximise sample pick-up. The swab should be held at a 30° angle to the contact surface. With the same swab, repeat this process at right angles to the first streaks to ensure that the entire sample area is swabbed. Replace swab into transport tube ensuring that swab comes into contact with the transport medium by pushing down hard. Clean the surface tested with disinfectant.
", "position": 1, "completed": false, "completed_on": null, "user_id": 202, "created_at": "2018-11-15T09:56:30.480Z", "updated_at": "2019-02-19T14:13:06.622Z", "last_modified_by_id": 202, "protocol_id": 3544 }, "checklists": [ { "checklist": { "id": 809, "name": "Sample locations:", "step_id": 4521, "created_at": "2018-11-15T09:56:30.495Z", "updated_at": "2018-11-15T09:56:30.495Z", "created_by_id": 202, "last_modified_by_id": 202 }, "checklist_items": [ { "id": 3405, "text": "Filling needle", "checked": false, "checklist_id": 809, "created_at": "2018-11-15T09:56:30.509Z", "updated_at": "2018-11-15T10:08:47.925Z", "created_by_id": 202, "last_modified_by_id": 202, "position": 0 }, { "id": 3406, "text": "Interior of the stopper chute", "checked": false, "checklist_id": 809, "created_at": "2018-11-15T09:56:30.522Z", "updated_at": "2018-11-15T10:08:47.935Z", "created_by_id": 202, "last_modified_by_id": 202, "position": 1 }, { "id": 3407, "text": "Helix/in-fed worm", "checked": false, "checklist_id": 809, "created_at": "2018-11-15T09:56:30.534Z", "updated_at": "2018-11-15T10:08:47.944Z", "created_by_id": 202, "last_modified_by_id": 202, "position": 2 }, { "id": 3408, "text": "In-feed belt", "checked": false, "checklist_id": 809, "created_at": "2018-11-15T09:56:30.546Z", "updated_at": "2018-11-15T10:08:47.954Z", "created_by_id": 202, "last_modified_by_id": 202, "position": 3 } ] } ], "step_comments": [], "step_assets": [], "assets": [], "step_tables": [], "tables": [] }, { "step": { "id": 5172, "name": "Plating of swabs", "description": "\nFollow the guidelines below for plating of swabs.
", "position": 4, "completed": false, "completed_on": null, "user_id": 202, "created_at": "2018-12-12T11:02:15.588Z", "updated_at": "2019-01-25T12:15:21.953Z", "last_modified_by_id": 202, "protocol_id": 3544 }, "checklists": [ { "checklist": { "id": 952, "name": "Guidelines:", "step_id": 5172, "created_at": "2018-12-21T06:58:27.768Z", "updated_at": "2018-12-21T06:58:27.768Z", "created_by_id": null, "last_modified_by_id": null }, "checklist_items": [ { "id": 3906, "text": "Prepare a safe and sterile workspace: Sterilize all instruments, solutions, and media prior to using them for plating procedures. Clear away all materials cluttering your work area on the laboratory bench. Clean work area with disinfectant to minimize possible contamination.", "checked": false, "checklist_id": 952, "created_at": "2018-12-21T06:58:27.770Z", "updated_at": "2018-12-21T06:58:27.770Z", "created_by_id": null, "last_modified_by_id": null, "position": 0 }, { "id": 3907, "text": "Set up a Bunsen burner: Work slowly, carefully, and deliberately within the sterile field area created by the updraft of the flame.", "checked": false, "checklist_id": 952, "created_at": "2018-12-21T06:58:27.777Z", "updated_at": "2018-12-21T06:58:27.777Z", "created_by_id": null, "last_modified_by_id": null, "position": 1 }, { "id": 3908, "text": "Prepare lab equipment and settle plates.", "checked": false, "checklist_id": 952, "created_at": "2018-12-21T06:58:27.784Z", "updated_at": "2018-12-21T06:58:27.784Z", "created_by_id": null, "last_modified_by_id": null, "position": 2 }, { "id": 3909, "text": "Arrange all the supplies needed for the procedure on the laboratory bench near the sterile field. Make sure all the materials are properly labelled.", "checked": false, "checklist_id": 952, "created_at": "2018-12-21T06:58:27.791Z", "updated_at": "2018-12-21T06:58:27.791Z", "created_by_id": null, "last_modified_by_id": null, "position": 3 }, { "id": 3910, "text": "Carefully open the lid of the plate. A standard streaking out method should be used when plating out the swab. The method should ensure that the swab is rotated as it is run over the surface of the media to ensure that any microorganisms recovered from the surface sample are deposited onto the surface of the plate.\r\nReplace the lid.", "checked": false, "checklist_id": 952, "created_at": "2018-12-21T06:58:27.797Z", "updated_at": "2018-12-21T06:58:27.797Z", "created_by_id": null, "last_modified_by_id": null, "position": 4 }, { "id": 3911, "text": "Complete and enclose all the necessary documentation.", "checked": false, "checklist_id": 952, "created_at": "2018-12-21T06:58:27.804Z", "updated_at": "2018-12-21T06:58:27.804Z", "created_by_id": null, "last_modified_by_id": null, "position": 5 } ] } ], "step_comments": [], "step_assets": [], "assets": [], "step_tables": [], "tables": [] }, { "step": { "id": 4519, "name": "Preparation of swabs before sampling", "description": "\nMake sure the sampling surface it is dry. Assemble the needle and 10 mL syringe. Open the ampoule of 0.9% NaCl Injection BP and draw up the contents. Push a small amount of liquid (about 0.5 mL) from the syringe directly onto the swab. Do not allow the needle tip to come into contact with the swab.
", "position": 0, "completed": false, "completed_on": null, "user_id": 202, "created_at": "2018-11-15T09:56:30.420Z", "updated_at": "2019-01-29T12:36:12.805Z", "last_modified_by_id": 202, "protocol_id": 3544 }, "checklists": [], "step_comments": [], "step_assets": [], "assets": [], "step_tables": [], "tables": [] } ] } ], "results": [] }, { "my_module": { "id": 2195, "name": "Incubation", "due_date": null, "description": "Incubation instructions for all types of plates", "x": 69, "y": 16, "my_module_group_id": 1250, "created_at": "2018-11-15T09:56:32.117Z", "updated_at": "2019-02-19T14:12:22.372Z", "archived": false, "archived_on": null, "created_by_id": 202, "last_modified_by_id": null, "archived_by_id": null, "restored_by_id": null, "restored_on": null, "workflow_order": 4, "experiment_id": 440, "state": "uncompleted", "completed_on": null }, "outputs": [ { "id": 6281, "input_id": 2196, "output_id": 2195 }, { "id": 6282, "input_id": 2200, "output_id": 2195 } ], "my_module_tags": [], "task_comments": [], "my_module_repository_rows": [], "user_my_modules": [], "protocols": [ { "protocol": { "id": 3554, "name": null, "authors": null, "description": null, "added_by_id": null, "my_module_id": 2195, "team_id": 1, "protocol_type": "unlinked", "parent_id": null, "parent_updated_at": null, "archived_by_id": null, "archived_on": null, "restored_by_id": null, "restored_on": null, "created_at": "2018-11-15T09:56:32.169Z", "updated_at": "2019-02-20T07:55:58.620Z", "published_on": null, "nr_of_linked_children": 0 }, "protocol_protocol_keywords": [], "steps": [ { "step": { "id": 4553, "name": "Incubation for growing bacteria", "description": "\n\nImportant!: If the medium is dropped or touched by a person performing the sampling then this should be reported, the sample should be marked accordingly and treated as usual. Under no circumstances should samples that have been taken be refrigerated.Follow the guidelines:
", "position": 1, "completed": false, "completed_on": null, "user_id": 202, "created_at": "2019-02-04T13:21:21.550Z", "updated_at": "2019-02-04T13:30:56.251Z", "last_modified_by_id": 202, "protocol_id": 4196 }, "checklists": [ { "checklist": { "id": 1014, "name": "Guidelines:", "step_id": 5496, "created_at": "2019-02-04T13:30:56.249Z", "updated_at": "2019-02-04T13:30:56.249Z", "created_by_id": null, "last_modified_by_id": null }, "checklist_items": [ { "id": 4149, "text": "Add 4 to 5 drops of 3% H2O2 to in a test tube.", "checked": false, "checklist_id": 1014, "created_at": "2019-02-04T13:30:56.251Z", "updated_at": "2019-02-04T13:30:56.251Z", "created_by_id": null, "last_modified_by_id": null, "position": 0 }, { "id": 4150, "text": "Using a wooden applicator stick, collect a small amount of organism from a well-isolated 18- to 24-hour colony and place into the test tube (Note: Be careful not to pick up any agar).", "checked": false, "checklist_id": 1014, "created_at": "2019-02-04T13:30:56.260Z", "updated_at": "2019-02-04T13:30:56.260Z", "created_by_id": null, "last_modified_by_id": null, "position": 1 }, { "id": 4151, "text": "Place the tube against a dark background and observe for immediate bubble formation at the end of the wooden applicator stick.", "checked": false, "checklist_id": 1014, "created_at": "2019-02-04T13:30:56.267Z", "updated_at": "2019-02-04T13:30:56.267Z", "created_by_id": null, "last_modified_by_id": null, "position": 2 } ] } ], "step_comments": [], "step_assets": [], "assets": [], "step_tables": [], "tables": [] }, { "step": { "id": 5495, "name": "Slide test", "description": "\nFollow the guidelines bellow.
", "position": 0, "completed": false, "completed_on": null, "user_id": 202, "created_at": "2019-02-04T13:18:55.135Z", "updated_at": "2019-02-04T13:18:55.140Z", "last_modified_by_id": 202, "protocol_id": 4196 }, "checklists": [ { "checklist": { "id": 1013, "name": "Guidelines:", "step_id": 5495, "created_at": "2019-02-04T13:18:55.138Z", "updated_at": "2019-02-04T13:18:55.138Z", "created_by_id": null, "last_modified_by_id": null }, "checklist_items": [ { "id": 4145, "text": "Transfer a small amount of bacterial colony to a surface of clean, dry glass slide using a loop", "checked": false, "checklist_id": 1013, "created_at": "2019-02-04T13:18:55.140Z", "updated_at": "2019-02-04T13:18:55.140Z", "created_by_id": null, "last_modified_by_id": null, "position": 0 }, { "id": 4146, "text": "Place a drop of 3% H2O2 on to the slide and mix.", "checked": false, "checklist_id": 1013, "created_at": "2019-02-04T13:18:55.155Z", "updated_at": "2019-02-04T13:18:55.155Z", "created_by_id": null, "last_modified_by_id": null, "position": 1 }, { "id": 4147, "text": "Look for a rapid evolution of oxygen (within 5-10 seconds) as evidenced by bubbling.", "checked": false, "checklist_id": 1013, "created_at": "2019-02-04T13:18:55.163Z", "updated_at": "2019-02-04T13:18:55.163Z", "created_by_id": null, "last_modified_by_id": null, "position": 2 }, { "id": 4148, "text": "Dispose of your slide in the biohazard glass disposal container.", "checked": false, "checklist_id": 1013, "created_at": "2019-02-04T13:21:32.881Z", "updated_at": "2019-02-04T13:21:32.881Z", "created_by_id": null, "last_modified_by_id": null, "position": 3 } ] } ], "step_comments": [], "step_assets": [], "assets": [], "step_tables": [], "tables": [] }, { "step": { "id": 5497, "name": "Results interpretation", "description": "\n\nCatalase is an enzyme, which is produced by microorganisms that live in oxygenated environments to neutralize toxic forms of oxygen metabolites. Anaerobes generally lack the catalase enzyme. Catalase enzyme hydrolyzes the hydrogen peroxide and forms bubbles.Follow the guidelines bellow:
", "position": 0, "completed": false, "completed_on": null, "user_id": 202, "created_at": "2019-02-01T14:49:04.449Z", "updated_at": "2019-02-01T14:49:04.453Z", "last_modified_by_id": 202, "protocol_id": 4195 }, "checklists": [ { "checklist": { "id": 1012, "name": "Guidelines:", "step_id": 5492, "created_at": "2019-02-01T14:49:04.452Z", "updated_at": "2019-02-01T14:49:04.452Z", "created_by_id": null, "last_modified_by_id": null }, "checklist_items": [ { "id": 4141, "text": "Take a filter paper soaked with the substrate tetramethyl-p-phenylenediamine dihydrochloride.", "checked": false, "checklist_id": 1012, "created_at": "2019-02-01T14:49:04.454Z", "updated_at": "2019-02-01T14:49:04.454Z", "created_by_id": null, "last_modified_by_id": null, "position": 0 }, { "id": 4142, "text": "Moisten the paper with a sterile distilled water.", "checked": false, "checklist_id": 1012, "created_at": "2019-02-01T14:49:04.462Z", "updated_at": "2019-02-01T14:49:04.462Z", "created_by_id": null, "last_modified_by_id": null, "position": 1 }, { "id": 4143, "text": "Pick the colony to be tested with wooden or platinum loop and smear in the filter paper.", "checked": false, "checklist_id": 1012, "created_at": "2019-02-01T14:49:04.469Z", "updated_at": "2019-02-01T14:49:04.469Z", "created_by_id": null, "last_modified_by_id": null, "position": 2 }, { "id": 4144, "text": "Observe inoculated area of paper for a color change to deep blue or purple within 10-30 seconds.", "checked": false, "checklist_id": 1012, "created_at": "2019-02-01T14:49:04.475Z", "updated_at": "2019-02-01T14:49:04.475Z", "created_by_id": null, "last_modified_by_id": null, "position": 3 } ] } ], "step_comments": [], "step_assets": [], "assets": [], "step_tables": [], "tables": [] }, { "step": { "id": 5493, "name": "Results interpretation", "description": "\nOxidase test is used for identification of Pseudomonas spp, Aeromonas spp, Neisserias spp, Vibrio spp and Pasteurella spp, all of which produces oxidase enzyme.
Blue-purple colour: Oxidase positive
No purple colour: Oxidase negative
Wipe the slides with spirit or alcohol. After cleaning, dry the slides and place them on laboratory towels until ready for use. Drawing a circle on the underside of the slide using a glassware-marking pen may be helpful to clearly designate the area in which you will prepare the smear. You may also label the slide with the initials of the name of the organism on the edge of the slide.
With a sterile cooled loop, place a drop of sterile water or saline solution on the slide. Sterilize and cool the loop again and pick up a very small sample of a bacterial colony and gently stir into the drop of water/saline on the slide to create an emulsion.
Heat fixing kills the bacteria in the smear, firmly adheres the smear to the slide, and allows the sample to more readily take up stains.
Allow the smear to air dry. After the smear has air-dried, hold the slide at one end and pass the entire slide through the flame of a Bunsen burner two to three times with the smear-side up.
Follow the guidelines below:
Important: Great care must be taken when heating the carbol fuchsin especially if staining is carried out over a tray or other container in which highly flammable chemicals have collected from the previous staining. Only a small flame should be applied under the slides using an ignited swab previously dampened with a few drops of acid alcohol or 70% v/v ethanol or methanol. Do not use a large ethanol soaked swab because this is a fire risk.
Follow the guidelines below.
", "position": 3, "completed": false, "completed_on": null, "user_id": 202, "created_at": "2019-02-01T14:18:22.722Z", "updated_at": "2019-02-20T07:35:23.931Z", "last_modified_by_id": 202, "protocol_id": 4193 }, "checklists": [ { "checklist": { "id": 1010, "name": "Guidelines:", "step_id": 5488, "created_at": "2019-02-01T14:18:22.724Z", "updated_at": "2019-02-01T14:18:22.724Z", "created_by_id": null, "last_modified_by_id": null }, "checklist_items": [ { "id": 4124, "text": "Flood air-dried, heat-fixed smear of cells for 1 minute with crystal violet staining reagent. Please note that the quality of the smear (too heavy or too light cell concentration) will affect the Gram Stain results.", "checked": false, "checklist_id": 1010, "created_at": "2019-02-01T14:18:22.727Z", "updated_at": "2019-02-01T14:18:22.727Z", "created_by_id": null, "last_modified_by_id": null, "position": 0 }, { "id": 4125, "text": "Wash slide in a gentle and indirect stream of tap water for 2 seconds.", "checked": false, "checklist_id": 1010, "created_at": "2019-02-01T14:18:22.742Z", "updated_at": "2019-02-01T14:18:22.742Z", "created_by_id": null, "last_modified_by_id": null, "position": 1 }, { "id": 4126, "text": "Flood slide with the mordant: Gram's iodine. Wait 1 minute.", "checked": false, "checklist_id": 1010, "created_at": "2019-02-01T14:18:22.753Z", "updated_at": "2019-02-01T14:18:22.753Z", "created_by_id": null, "last_modified_by_id": null, "position": 2 }, { "id": 4127, "text": "Wash slide in a gentle and indirect stream of tap water for 2 seconds.", "checked": false, "checklist_id": 1010, "created_at": "2019-02-01T14:18:22.764Z", "updated_at": "2019-02-01T14:18:22.764Z", "created_by_id": null, "last_modified_by_id": null, "position": 3 }, { "id": 4128, "text": "Flood slide with decolorizing agent. Wait 15 seconds or add drop by drop to slide until decolorizing agent running from the slide runs clear.", "checked": false, "checklist_id": 1010, "created_at": "2019-02-01T14:18:22.771Z", "updated_at": "2019-02-01T14:18:22.771Z", "created_by_id": null, "last_modified_by_id": null, "position": 4 }, { "id": 4129, "text": "Flood slide with counterstain, safranin. Wait 30 seconds to 1 minute.", "checked": false, "checklist_id": 1010, "created_at": "2019-02-01T14:18:22.779Z", "updated_at": "2019-02-01T14:18:22.779Z", "created_by_id": null, "last_modified_by_id": null, "position": 5 }, { "id": 4130, "text": "Wash slide in a gentile and indirect stream of tap water until no color appears in the effluent and then blot dry with absorbent paper.", "checked": false, "checklist_id": 1010, "created_at": "2019-02-01T14:18:22.786Z", "updated_at": "2019-02-01T14:18:22.786Z", "created_by_id": null, "last_modified_by_id": null, "position": 6 }, { "id": 4131, "text": "Observe the results of the staining procedure under oil immersion using a Brightfield microscope. At the completion of the Gram Stain, gram-negative bacteria will stain pink/red and gram-positive bacteria will stain blue/purple.", "checked": false, "checklist_id": 1010, "created_at": "2019-02-01T14:18:22.793Z", "updated_at": "2019-02-01T14:18:22.793Z", "created_by_id": null, "last_modified_by_id": null, "position": 7 } ] } ], "step_comments": [], "step_assets": [ { "id": 3081, "step_id": 5488, "asset_id": 3743 } ], "assets": [ { "asset": { "id": 3743, "created_at": "2019-02-20T07:34:52.023Z", "updated_at": "2019-02-20T07:35:23.920Z", "created_by_id": 202, "last_modified_by_id": null, "estimated_size": 21906, "lock": null, "lock_ttl": null, "version": 1, "file_processing": false, "team_id": 1 }, "asset_blob": { "filename": "Gram_Stain_Procedure.jpg" } } ], "step_tables": [], "tables": [] }, { "step": { "id": 5475, "name": "Reagents", "description": "\n\nReagents bellow can be made or purchased.
Crystal Violet Staining Reagent:
Solution A for crystal violet staining reagent
Working Solution:
\nWipe the slides with spirit or alcohol. After cleaning, dry the slides and place them on laboratory towels until ready for use. Drawing a circle on the underside of the slide using a glassware-marking pen may be helpful to clearly designate the area in which you will prepare the smear. You may also label the slide with the initials of the name of the organism on the edge of the slide.
With a sterile cooled loop, place a drop of sterile water or saline solution on the slide. Sterilize and cool the loop again and pick up a very small sample of a bacterial colony and gently stir into the drop of water/saline on the slide to create an emulsion.
Heat fixing kills the bacteria in the smear, firmly adheres the smear to the slide, and allows the sample to more readily take up stains.
Allow the smear to air dry. After the smear has air-dried, hold the slide at one end and pass the entire slide through the flame of a Bunsen burner two to three times with the smear-side up.
\n
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After all the samples are taken, collect all sample plates together, remove them from the sampling area and incubate as directed. Complete and enclose the necessary documentation [#Counting and recording...~tsk~ZU].
", "position": 3, "completed": false, "completed_on": null, "user_id": 202, "created_at": "2018-11-15T09:56:29.770Z", "updated_at": "2019-01-25T12:42:42.158Z", "last_modified_by_id": 202, "protocol_id": 3540 }, "checklists": [], "step_comments": [], "step_assets": [], "assets": [], "step_tables": [], "tables": [] }, { "step": { "id": 5449, "name": "Labeling of plates", "description": "\n\nAssemble the plates required and label them. Ensure that correct information about sampling is written on the base of the plate with a permanent ink marker.
The following sampling details should be recorded on the plate:
Preparation of sampling location and air sampler: Ensure air sampler is fully charged. Sanitize the external surfaces of the unit with sterile 70% isopropyl alcohol. Press the black colour button on the bottom side to start the air sampler.
Sampling: Select standard mode and press confirm. Select 10 000L of air volume and press enter key to confirm. After pressing the enter key START FOR 10 000 will appear on display. Carefully remove the cover from the sieve. Place the settle plate on the air sampler and screw the sieve head. Start the air sampler by pressing enter. Sampling should last around 10 minutes.
After air sampling is finished: Unscrew the sieve head of the air sampler. Carefully remove the agar plate and cover it with lid. Swab areas where plates have been exposed with a suitable disinfectant to remove any trace of media or condensation from the lids, which may contaminate the room. Swab areas where plates have been exposed with a suitable disinfectant to remove any trace of media or condensation from the lids, which may contaminate the clean room.
All samples should be discarded according to procedures.
", "position": 4, "completed": false, "completed_on": null, "user_id": 202, "created_at": "2018-12-12T11:42:25.451Z", "updated_at": "2018-12-12T11:42:25.451Z", "last_modified_by_id": 202, "protocol_id": 3556 }, "checklists": [], "step_comments": [], "step_assets": [], "assets": [], "step_tables": [], "tables": [] }, { "step": { "id": 4558, "name": "Streak plate procedure", "description": "\nFollow streaking procedure as displayed on image and guidelines below.
", "position": 1, "completed": false, "completed_on": null, "user_id": 202, "created_at": "2018-11-15T09:56:32.459Z", "updated_at": "2019-02-20T07:34:22.994Z", "last_modified_by_id": 202, "protocol_id": 3556 }, "checklists": [ { "checklist": { "id": 953, "name": "Guidelines:", "step_id": 4558, "created_at": "2018-12-21T07:19:34.041Z", "updated_at": "2018-12-21T07:19:34.041Z", "created_by_id": null, "last_modified_by_id": null }, "checklist_items": [ { "id": 3912, "text": "Label the base of the plate. Open the lids and flame metal loop using a Bunsen burner before obtaining the inoculum for the plate.", "checked": false, "checklist_id": 953, "created_at": "2018-12-21T07:19:34.043Z", "updated_at": "2018-12-21T07:19:34.043Z", "created_by_id": null, "last_modified_by_id": null, "position": 0 }, { "id": 3913, "text": "The metal should become red hot. Move the wire so the flame approaches the loop. Spread sample over about one-quarter of the surface of the medium using a rapid, smooth, back-and-forth motion.", "checked": false, "checklist_id": 953, "created_at": "2018-12-21T07:19:34.053Z", "updated_at": "2018-12-21T07:19:34.053Z", "created_by_id": null, "last_modified_by_id": null, "position": 1 }, { "id": 3914, "text": "Lift the bottom half of an inverted plate from the bench. Move the loop back and forth many times across the agar surface from the rim to the centre of the plate. Reflame the metal loop. Turn the Petri dish 90 ° to streak the second quadrant. Using the back-and-forth pattern, cross over the last half of the streaks in the first quadrant then move into the empty second quadrant. Repeat until all 4 quadrants are done.", "checked": false, "checklist_id": 953, "created_at": "2018-12-21T07:19:34.060Z", "updated_at": "2018-12-21T07:19:34.060Z", "created_by_id": null, "last_modified_by_id": null, "position": 2 }, { "id": 3915, "text": "Cover all the plates with their lids.", "checked": false, "checklist_id": 953, "created_at": "2018-12-21T07:19:34.067Z", "updated_at": "2018-12-21T07:19:34.067Z", "created_by_id": null, "last_modified_by_id": null, "position": 3 }, { "id": 3916, "text": "Clean the area with disinfectant.", "checked": false, "checklist_id": 953, "created_at": "2018-12-21T07:19:34.073Z", "updated_at": "2018-12-21T07:19:34.073Z", "created_by_id": null, "last_modified_by_id": null, "position": 4 } ] } ], "step_comments": [], "step_assets": [ { "id": 3079, "step_id": 4558, "asset_id": 3741 } ], "assets": [ { "asset": { "id": 3741, "created_at": "2019-02-20T07:33:40.415Z", "updated_at": "2019-02-20T07:34:22.986Z", "created_by_id": 202, "last_modified_by_id": null, "estimated_size": 48768, "lock": null, "lock_ttl": null, "version": 1, "file_processing": false, "team_id": 1 }, "asset_blob": { "filename": "Plate_Streaking.jpg" } } ], "step_tables": [], "tables": [] }, { "step": { "id": 5175, "name": "Incubation", "description": "\nFollow the protocol for incubation in [#Incubation~tsk~ZP]
", "position": 2, "completed": false, "completed_on": null, "user_id": 202, "created_at": "2018-12-12T11:36:16.853Z", "updated_at": "2019-02-19T14:27:45.703Z", "last_modified_by_id": 202, "protocol_id": 3556 }, "checklists": [], "step_comments": [], "step_assets": [], "assets": [], "step_tables": [], "tables": [] }, { "step": { "id": 4559, "name": "Prepare a safe and sterile workspace", "description": "\nUse different tests to identify microorganisms:
\nAfter all the samples are taken, collect all sample plates together, remove them from the sampling area and incubate as directed. Complete and enclose the necessary documentation [#Counting and recording...~tsk~ZU].
", "position": 3, "completed": false, "completed_on": null, "user_id": 202, "created_at": "2018-11-15T09:56:31.742Z", "updated_at": "2019-01-25T12:41:23.609Z", "last_modified_by_id": 202, "protocol_id": 3552 }, "checklists": [], "step_comments": [], "step_assets": [], "assets": [], "step_tables": [], "tables": [] }, { "step": { "id": 5447, "name": "Labeling of plates", "description": "\n\nAssemble the plates required and label them. Ensure that correct information about sampling is written on the base of the plate with a permanent ink marker.
The following sampling details should be recorded on the plate:
Transfer the contact plates into the area to be tested. Enter the area according to the procedures. Ensure that the testing area is dry.
When sampling surfaces with contact plates, remove the lid and place the agar surface in maximum contact with the sampling site for 10 seconds by applying a constant force spread evenly over the whole contact plate without twisting or sliding and avoiding the creation of bubbles. After contact with the sample surface, remove and replace the lid, avoiding any unnecessary hand movement near or over the agar surface. Clean it with disinfectant to remove any possible traces of agar.
Testing conditions: Sampling should take place at the end of a work session, but before the person would carry out any cleaning. Before performing the test the person should ensure that gloves are dry and free of any disinfectant.
Glove test: The person doing the sampling should lift the lid of the plate, with the opposite hand to that being tested, and keeping hold of the lid in the other hand, touch the agar surface with the tips of all fingers then the thumb (in the gap on the plate behind where fingers were tested) on the hand being tested. A firm and even pressure should be applied for approximately 5 to 10 seconds, taking care not to damage the agar surface. Replace the lid of the plate. Repeat for the other hand. Ensure that the glove surfaces tested are cleaned with a suitable disinfectant to remove any possible traces of agar before performing any other operations.
Gowning test: The person doing the sampling should lift the lid of the plate and keep hold of the lid, touch the gown in the chest and clavicle area with agar surface. A firm and even pressure should be applied for approximately 5 to 10 seconds, taking care not to damage the agar surface. Replace the lid of the plate. Ensure that the gown surfaces tested are cleaned with a suitable disinfectant to remove any possible traces of agar before performing any other operations.