Isolasi bakteri dengan teknik pengenceran dan penanaman secara spread plate

Anriani Puspita

10 May 202206:23

Summary

TLDRThis video tutorial demonstrates the process of bacterial isolation using serial dilution and spread plate techniques. It begins with preparing tools and materials, including urine samples, dilution tubes, and sterile equipment. The urine sample is serially diluted, mixed with a vortex mixer, and then plated onto agar media using a spreader. The spreader is sterilized with alcohol and flame before use. The final step involves incubating the plates at the appropriate temperature to grow bacterial colonies.

Takeaways

🔬 The video demonstrates a method for bacterial isolation using serial dilution and spread plate techniques.
💧 The sample used in the demonstration is urine.
🧪 Preparation includes sterilized equipment and materials such as a Bunsen burner, alcohol sprayer, and a Vortex mixer.
🔥 Aseptic technique is maintained by sterilizing the workspace with 70% alcohol and using a Bunsen burner flame.
📦 The process starts with taking 1 mL of urine sample using a micropipette.
🌡️ The sample is then transferred to a dilution tube containing 9 mL of sterile water, creating a 10-fold dilution.
🔁 The dilution tube is homogenized using a Vortex mixer to ensure even distribution of the sample.
🔄 This dilution process is repeated through a series of tubes to achieve higher dilutions.
📐 The final dilution is plated onto an agar medium using a spreader, previously sterilized with 70% alcohol and a Bunsen burner.
🌡️ The spreader is allowed to cool before use and is sterilized again after use.
🌱 The petri dishes are incubated at the appropriate temperature to allow for bacterial growth.

Q & A

What is the main topic of the video?
-The main topic of the video is the isolation of bacteria using a technique called serial dilution and the spread plate method.
What is the first sample used in the video?
-The first sample used in the video is urine.
What are the materials needed for the serial dilution process as mentioned in the video?
-The materials needed include sterile water, a pipette, a Vortex mixer, a Bunsen burner, alcohol spray, and a disposable loop.
What is the purpose of using a Bunsen burner in the process?
-The Bunsen burner is used to flame the disposable loop and other tools to sterilize them and maintain aseptic conditions during the process.
How much urine sample is taken initially for the process?
-One milliliter (1 mL) of urine sample is taken initially using a micropipette.
What is the purpose of adding the urine sample to the first dilution tube containing sterile water?
-The urine sample is added to the first dilution tube to create a 1:10 dilution, which helps in isolating and counting the bacteria present in the sample.
How is the mixture homogenized after adding the urine sample to the dilution tube?
-The mixture is homogenized using a Vortex mixer to ensure the sample is evenly distributed in the dilution medium.
What is the next step after homogenization in the serial dilution process?
-The next step is to take 1 mL of the homogenized mixture and transfer it to the next dilution tube containing 9 mL of sterile water, and then repeat the homogenization process.
How many dilution tubes are used in the process, and what is the final step after the last dilution?
-Four dilution tubes are used in the process. After the last dilution, the final step is to perform the spread plate method by taking 0.1 mL of the last dilution and spreading it onto an agar plate.
What is the purpose of sterilizing the spreader before spreading the sample on the agar plate?
-The spreader is sterilized by flaming it over a Bunsen burner to prevent contamination of the agar plate with unwanted microorganisms.
What is the final step after spreading the sample on the agar plate?
-The final step is to incubate the petri dishes containing the spread plates at the appropriate temperature to allow the bacteria to grow.

Outlines

00:00

🔬 Bacterial Isolation Process

This paragraph describes a method for bacterial isolation using serial dilution and spread plate techniques. The process begins with preparing urine samples and various equipment including dilution tubes, sterile water, pipettes, a Bunsen burner, a Vortex mixer, and an alcohol sprayer. The workspace is sanitized with 70% alcohol, and a Bunsen burner is used to maintain aseptic conditions. Urine samples are taken and diluted in a series of dilution tubes, each containing sterile water, and mixed using a Vortex mixer. The dilution process is repeated until the fourth dilution tube. The final step involves spreading 0.1 mL of the last dilution onto an agar plate for incubation.

05:00

🌡️ Spread Plate Technique and Incubation

This paragraph details the spread plate technique for evenly distributing the bacterial sample on an agar medium. A sterile spreader, or drugalsky, is used after being cooled and sterilized with 70% alcohol and heated over a Bunsen burner. The spreader is used to evenly distribute the 0.1 mL of bacterial solution on the agar plate. After use, the spreader is again heated to sterilize it before storage. The final step is to cover the petri dish and incubate it at the appropriate temperature to allow bacterial growth. The video concludes with a thank you note, hoping the information is useful.

Mindmap

Keywords

💡Isolation

Isolation in the context of microbiology refers to the process of separating a specific type of microorganism from a mixture of different organisms. In the video, isolation is the main goal, as it describes the method to separate bacteria from a urine sample. The process involves serial dilution and plating techniques to grow and identify individual bacterial colonies.

💡Serial Dilution

Serial dilution is a technique used to reduce the concentration of a solution in a series of steps. In the video, serial dilution is mentioned as a method to prepare the urine sample by gradually diluting it with sterile water, which helps in isolating bacteria at different concentration levels.

💡Spread Plate

Spread plating is a method of inoculating a culture medium with a diluted sample by spreading it across the surface of the medium. This technique is used to grow isolated colonies of bacteria for further study. The video script describes the use of this method to plate the diluted urine sample onto an agar medium.

💡Urine Sample

A urine sample is a biological specimen collected from the human body for diagnostic purposes. In the video, urine is the source material from which bacteria are to be isolated. The script mentions the preparation of the urine sample as the starting point for the isolation process.

💡Sterile Aquadest

Sterile aquadest, or distilled water, is used in microbiology to dilute samples and prepare media without introducing contaminants. The script refers to the use of sterile aquadest to dilute the urine sample during the serial dilution process.

💡Micropipette

A micropipette is a tool used to transfer small volumes of liquid with precision. In the video, a micropipette is used to measure and transfer urine samples and dilutions, ensuring accurate volumes for the isolation process.

💡Vortex Mixer

A vortex mixer is a device that spins samples to mix them thoroughly. In the video, a vortex mixer is used to homogenize the urine sample after dilution, ensuring that the bacteria are evenly distributed within the solution.

💡Bunsen Burner

A Bunsen burner is a type of gas burner used for heating, sterilization, and flame applications in a laboratory. The script mentions using a Bunsen burner to sterilize tools like the spreader (drugalsky) and to maintain an aseptic environment during the procedure.

💡Agar Medium

Agar medium is a gelatinous substance used as a growth medium for microorganisms. In the video, agar is the medium onto which the diluted urine sample is spread to allow bacterial growth and isolation.

💡Inoculation

Inoculation is the process of introducing a microorganism into a culture medium. The script describes the inoculation of the agar medium with the diluted urine sample to grow bacterial colonies.

💡Incubation

Incubation is the process of maintaining optimal conditions, such as temperature and humidity, to allow the growth of microorganisms. The video concludes with the incubation of the inoculated agar plates to allow the bacteria to grow and form visible colonies.

Highlights

Introduction to bacterial isolation using gradient dilution and spread plate techniques.

Preparation of tools and materials including urine sample, dilution tubes, and agar media.

Use of Google Sky for media preparation.

Sterilization of equipment and workspace with 70% alcohol.

Ignition of Bunsen burner for aseptic work.

Preparation of urine sample using a micropipette.

Transfer of urine sample to the first dilution tube containing sterile water.

Homogenization of the sample using a Vortex mixer.

Sequential dilution process using new pipettes for each step.

Transfer of diluted sample to subsequent dilution tubes.

Final dilution step before plating.

Use of spread plate method for sample plating.

Sterilization of the spreader (drugalsky) with 70% alcohol and Bunsen burner.

Spreading of 0.1 mL of the final dilution onto the agar medium.

Even distribution of the sample on the medium using a cooled spreader.

Re-sterilization and storage of the spreader after use.

Incubation of the petri dishes at the appropriate temperature.

Conclusion and expression of gratitude for the tutorial.