It is a device utilized for the pressure measurement of gases or liquids. Pressure is an expression of the force necessary to stop a fluid from expanding.
A pressure sensor acts like a transducer as it will generate a signal as a function of the pressure imposed.
These devices can vary drastically in technology, design, performance, application, and cost.
They can be alternately termed as pressure transducers or even something such as manometers.
Pressure sensors are used for control and monitoring in thousands of everyday applications. Pressure sensors can also be used to indirectly measure other variables such as fluid/gas flow, speed, water level, and altitude.
For budding gynecologists, the path to be an IVF specialist is full of challenges. With an increase in the number of clinics, information pertaining to IVF is available but it is still difficult for an outsider to peep in and grasp the enormity of it. There also may be a lack of clarity on how the process works and many times, prospective clinicians or embryologists need to get acquainted with the basics, so that they can choose IVF as an option.
EART (Embryology Academy for Research and Training) held an introductory session on February 13, 2019, for doctors who may be a part of our The doctors (about 25 in number) enrolled for the fellowship at the prestigious ‘21st Century’, Nadkarni Hospital (Vapi and Surat) participated enthusiastically and were able to gain a very thorough insight into the field of IVF.IVF sector in future and this orientation program laid the foundation for the same.
The sessions were conducted by the EART team and the event was judiciously divided into theoretical sessions, practical demonstrations, and a tour. Theoretical sessions in the form of talks were presented by Dr. Kersi Avari (EART Scientific Director) and Dr. Steve Lewitt (Director Clinical applications, Cooper Surgicals) who together enlightened the audience on embryology in general along with the emphasis on embryo transfer and ovum pick up. Mr. Ashish Modi (Managing Director of Shivani Scientific Industries Pvt. Ltd.) held a crucial talk on IVF scenario in India. Post lunch, practical demonstrations commenced which were divided into three workstations catering to different practical aspects such as ICSI, vitrification, and Andrology. (Conducted by Dr. Kersi Avari and Clinical Embryologists, Shridutt Gaitonde and Mohit Karkal)
The demonstrations covered everything but were short of hands-on owing to the paucity of time. A tour highlighting instrumentation and gadgets was done along with the sessions. From basics to advanced procedures such as ICSI, the participants were updated about the challenges and opportunities in the field of embryology, along with future avenues and clinical research.
The event organized on the premises of EART was highly interactive and the doctors were happy to be associated with the event which was a phenomenal success in terms of pushing the envelope in disseminating knowledge regarding IVF.
A magnetic stirrer or mixer is a laboratory device that employs a rotating magnetic field to cause a stir bar immersed in a liquid to spin quickly, thus stirring it!
The said magnetic field works either by rotating a magnet or using a set of stationary electromagnets which are placed beneath the vessel harboring the liquid.
They work well in glass vessels commonly used for chemical reactions as glass does not affect a magnetic field.
The limited size of the bars means that they can be used for small experiments, of says 4 liters or less. They also are difficult for usage in thick suspensions.
Magnetic stirrers are better than gear driven motorized stirrers as they are quieter and more efficient!
They often find applications in chemistry or biology and can be used inside hermetically closed vessels or systems. #LaboratoryInstrument #MagneticStirrer
A pH meter is an instrument which measures the hydrogen-ion activity in water-based solutions, indicating its acidity or alkalinity and is expressed as pH. The device measures the difference in electric potential between a pH electrode and a reference electrode and this difference relates to the acidity or pH of the solution.
pH meter is of many types and they range from simple or inexpensive pen-like devices to complex and expensive laboratory instruments with computer interfaces. Specialized forms are available for use in special applications such as harsh environments or the biological microenvironment.
Applications:
Chemical laboratory analysis and quality control is the most important application. pH meters are used for soil measurements in agriculture, determining water quality, brewing of wine, manufacturing, and healthcare or clinical applications including IVF units.
DNA MICROARRAY (also commonly known as biochips) is a collection of microscopic ‘DNA spots’ adhered to a solid surface which can be used for measuring the expression levels of a large number of genes simultaneously or to genotype multiple areas of a genome!
Each DNA spot contains a specific DNA sequence, also called as probes. These can be a short section of a gene or other DNA element which is used to hybridize a cDNA or cRNA sample (called target) under stringent conditions. Probe-target hybridization is detected and quantified by detection of fluorophore, silver or something called chemiluminescence- labeled targets to determine the relative presence of nucleic acid sequences in the target.
Many types of arrays exist and the broadest distinction is whether they are spatially arranged on a surface or on a coded bead.
DNA Microarray has many applications such as the identification of new genes along with knowing its expression or function, learning more about diseases and cancer, drug discovery and research.
A successful cryopreservation program has become the onus of fertility clinics all over. Importantly, as ‘freeze all’ cycles are slowly making inroads, the dependence on an effective cryopreservation program is the need of the hour.
Vitrification has been the choice lately for cryopreservation and can have slight variations depending upon the temperature, cryo devices and the brand of the media. Nevertheless, the basic principle and protocol are similar considering all these factors. Vitrification procedure is accomplished with the help of the available Vitrification media. The embryos are subjected to ES (Equilibration media) followed by its incorporation into VS (Vitrification media) before they are loaded on to a cryo device and plunged into liquid nitrogen. As an embryologist, I have always found Vitrification as more daunting to master than ICSI or other procedures. Unfortunately, there isn’t an exhaustive learning module with respect to the same. Fertility clinics do not follow a standard training program for Vitrification which may result into incompetency of the operator, poor survival rates and fluctuations in results!
The final step of embryo loading is the centerpiece of the whole procedure and the entire Vitrification program rests on the premise of that crucial last one minute. Naturally, the main focus will be on the last step which includes activities such as washing embryos in VS, loading them with minimal media on the cryo device and finally draining excessive media and plunging them into liquid nitrogen. The step is not difficult but certainly is challenging considering the microscopic aspects of the same coupled with the race with time.
The biggest challenge in Vitrification is the composite of speed with time! However, the technique can be effectively implemented after an extensive stepwise way to learn it by self devising a good training environment. The overall learning process may take longer to master but the initiation is critical.
Embryology Academy for Research and Trainingaffiliated with Shivani Scientific Industries Pvt. Ltd. has designed a two-day ‘Advanced Course in Cryopreservation’ where beginning to learn Vitrification can be made easy. The course offers both theoretical aspects and hands-on experience with different cryo devices and even mimicking an actual freezing procedure. EART provides materials such as microbeads to practice the loading step which gives the trainee a hang of the technique and ushers in confidence. Training with respect to liquid nitrogen handling also is given along with brochures and study material. Tips such as placement of the hand whilst loading, the importance of labeling the devices, overcoming the fears of washing embryos in VS, the hand cryo device and eye coordination hurdles and steps associated with thawing are meticulously covered. Information is also given in terms of how the cryo devices will be stored and how the nitrogen tanks will be managed or maintained.
Nevertheless, after this critical initiation, if you get to work in a fertility clinic, there is an extensive way to further master the Vitrification procedure. The following are the steps over the course of six months you can chart in order to gain proficiency in the same!
Firstly, when you get acquainted with the technique just learning the protocol isn’t the key. There has to be an understanding of the science behind it. I have seen people panicking when they see embryos shrinking in ES!
Secondly, for starters, I would reckon an embryologist to be familiar with all the components of a Vitrification system. The goal here would be to do things perfectly without time constraints. The last major step can be broken into mini steps for better acclimatization. He can learn to adjust the coarse knob of the microscope to pick up the embryos after they float owing to change in viscosity in VS, the focusing of the cryotop under the microscope, the placement of the hand, learning the process of washing and finally gaining the ability to load the embryos!
Thirdly, once you have perfected the step, time will come into the picture. Then the use of stopwatch and practicing loading in its required time becomes the focus. Initially, the loading time may exceed even two mins. With practice, slowly, you will reduce the necessary time and finally place yourself in the stipulated range of sixty to ninety seconds! This practice can be done with degenerated oocytes.
Fourthly, once you have taken care of the time, the next natural factor will be survival! This is the stage where you will use discarded embryos and actually freeze them as if you are performing an actual procedure. Here, even the ES will come into your practice and you will observe intricate details such as the reaction of the embryos with the media. At this stage, the thawing technique will also be incorporated as it too contributes in the overall survival. The results can be recorded and it will give an insight into the overall improvement of the operator.
Finally, once you are done with survival, it is always recommended to freeze for embryo donation patients as ED cycle harbors the best quality embryos. Blastocysts should be considered more in the initial phase of freezing which survives better than say D2/D3 embryos. This can be furthered into increasing the number of cryo devices progressively corresponding with checking your performance of the previously frozen cryo devices simultaneously.
This comprehensive way of learning Vitrification may take six months overall depending upon the cycles happening in a fertility clinic which certainly affects the availability of gametes and resources(such as expired media for practice). It can help attain better proficiency over the technique and the program can be robustly standardized in a working fertility center itself. Vitrification is a gateway for the embryos to hibernate in an ice age and maybe we can make their journey qualitatively smooth!
A ‘Flow Cytometer’ can analyze thousands of particles per second in real time and if configured as cell sorters, can actively separate particles with certain optical characteristics! Instead of producing a magnified image of the cell like a microscope, a Flow Cytometer offers high throughput, effective quantification of optical parameters on a cell by cell basis.
Based on the principle of flow cytometry, the simplest form of this instrument counts each cell as it passes the laser beam and records the level of fluorescence the cell emits; an attached computer generates plots of the number of cells as the ordinate and their fluorescence intensity as the abscissa. More sophisticated versions of the instrument are capable of sorting populations of cells into different containers according to their fluorescence profile.
A #flowcytometer has five main components: a flow cell, a measuring system, a detector, an amplification system, and a computer for analysis of the signals. Flow cytometry has many applications and is routinely used in basic research, clinical practice, and clinical trials.
Uses for flow cytometry include Cell counting – Cell sorting – Determining cell characteristics and function – Detecting microorganisms – Biomarker detection – Protein engineering detection – Diagnosis of health disorders such as blood cancers. #LaboratoryInstrument
Intra Cytoplasmic Sperm Injection has been the holy grail of an IVF lab. After its successful initiation in the early 1990s, the technique has had a massive impact on the industry. ICSI was all about tackling severe male infertility but now it is routinely used in a fertility clinic where ICSI has almost replaced IVF. Is too much of ICSI detrimental in the long run? Now, that is a matter of serious debate. However, ICSI is now one of the key techniques necessary for an embryologist to grow. It is a ticket to push you into seniority and mastering the same is a challenge.
But, unfortunately, in the wake of quickly acquiring this skill, budding embryologists try to fast track their learning process. This results in a half-baked approach which doesn’t help in gaining proficiency in ICSI.
I have self-trained myself in ICSI as I had joined a fertility clinic as a trainee. Resources initially were less but as the cases picked up, I got an opportunity to get myself acquainted with the process of imbibing the technique fully.
ICSI Procedure
For starters, ICSI is a procedure where a single sperm is injected into a mature oocyte with the help of a micromanipulator. At the face of it, the procedure may require a week to get basically trained but that may not prepare you to sail through the overwhelming tides of handling regular cases and unpredictable nature of the load of work. The following steps can help overcome pitfalls and result in an exhaustive preparedness into ICSI.
Know The ICSI Micromanipulator
Firstly, the fundamentals lie in knowing the machine and the micromanipulator or the microscope thoroughly. It is like you ought to marry the machine in order to get accustomed to every facet and aspect of its working. My experience has taught me that embryologists tend to bypass this essentiality. It is important to know about the condenser as much as it is important to know about the holding and the injecting pipette. Any type, whether it is RI or Narishige and even Eppendorf system, knowing intricate details of the machine will be the foundation on which you can even tackle troubleshooting issues.
Secondly, the alignment part is to be mastered. Here, I have seen people skipping certain steps that snowball into problems later when you are about to immobilize sperms or inject oocytes. Practicing alignment comes across as boring which makes people skip it or think it’s avoidable. This is the time when used needles should be incorporated and used to align as this will not only teach you to do it faster but also will make you realize needles are so sensitive to handle and they break! Details such as setting the angle and changing the filter with respect to objective get drowned in the midst of hurrying the alignment.
Preparing ICSI dish a priority
Thirdly, making an ICSI dish is also a priority. This will give you an idea of the pattern you will incorporate along with the number of sperms to be added in the PVP droplet. I have seen inexperience flowing when too many sperms are added in the PVP drop which becomes difficult for immobilization. Making a clean and clear dish along with adding an optimum amount of sperm is equally important as other steps.
Selection Of Sperm
Fourthly, sperm catching will follow. This step requires the maximum practice and attention as the morphology of chosen sperm is in your hands considering there won’t be a choice with respect to oocytes. Initially, the experience of sperm immobilization may be increasingly frustrating and this may lure trainees to jump into oocyte injection part which seems much easy and yes a bit glamorous. Basically, you have to train your eye to choose the correct sperm and the difficulty level must increase with Normozoospermia followed by severe OAT and even surgically retrieved samples. For beginners, the edge of a PVP sperm drop provides the necessary reservoir to begin practice sperm catching!
Oocyte Injection
Lastly, the Oocyte injection will be the final step. In this part, during training, you may not get actual oocytes and hence you may miss the actual feeling of injecting true mature eggs. The little suction to be applied for holding the egg, aspiration till you get a jerk like feeling, the observation of a funnel-shaped structure in the cytoplasm won’t be visible whilst injecting oocytes other than mature.
Once you have combined all the steps and are confident, then the process of trying to time and stimulating a real ICSI environment must begin. In this scenario, the injections of oocytes should commence in this order. First, it should be degenerated eggs followed by unfertilized or M1s or GVs. The latter may not indicate fertilization, but we can just check if they survive the injection process. This should be followed by giving two or three oocytes to inject from a patient exhibiting good quality oocytes and sperms. This should be followed by a progressive increase in a number of eggs with a corresponding increase in the level of difficulty. (say to TESA/PESA cases) Here, you will actually get exposed to different types and varieties of mature oocytes you have to handle. This should culminate in giving half the oocytes of the patient and then the whole patient under monitoring or supervision. A documentation of recording the fate of every mature egg injected (fertilization or degeneration) and the technique applied (smooth or rough) together will ensure in giving confidence in absorbing the said embryologist in routine ICSI procedures in the fertility clinic.
With the above steps, one should require about six months for doing ICSI routinely with respect to the availability of resources. ICSI is like learning to drive a car. The training at a motor school will teach you the basics but how you acclimatize to roads and different terrains will depend upon how you handle the car outside the school. It takes time before you get used to driving and so this above process to learn ICSI will do the same for this technique.
Tips on How to successfully tackle Vitrification In IVF Lab.
A successful cryopreservation program has become the onus of fertility clinics all over. Importantly, as ‘freeze all’ cycles are slowly making inroads, the dependence on an effective cryopreservation program is the need of the hour.
Vitrification has been the choice lately for cryopreservation and can have slight variations depending upon the temperature, cryo devices and the brand of the media. Nevertheless, the basic principle and protocol are similar considering all these factors. Vitrification procedure is accomplished with the help of the available Vitrification media. The embryos are subjected to ES (Equilibration media) followed by its incorporation into VS (Vitrification media) before they are loaded on to a cryo device and plunged into liquid nitrogen. As an embryologist, I have always found Vitrification as more daunting to master than ICSI or other procedures. Unfortunately, there isn’t an exhaustive learning module with respect to the same. Fertility clinics do not follow a standard training program for Vitrification which may result into incompetency of the operator, poor survival rates and fluctuations in results!
Vitrification of oocytes
The final step of embryo loading is the centerpiece of the whole procedure and the entire Vitrification program rests on the premise of that crucial last one minute. Naturally, the main focus will be on the last step which includes activities such as washing embryos in VS, loading them with minimal media on the cryo device and finally draining excessive media and plunging them into liquid nitrogen. The step is not difficult but certainly is meticulous considering the microscopic aspects of the same coupled with the race with time.
The biggest challenge in Vitrification is the composite of speed with time! However, the technique can be effectively implemented after an extensive stepwise way to learn it by self-devising a good training environment.
Vitrification has been the choice lately for cryopreservation
Firstly, when you get acquainted with the technique, just learning the protocol isn’t the key. There has to be an understanding of the science behind it. I have seen people panicking when they see embryos shrinking in ES!
Embryos Shrinking
Secondly, for starters, I would reckon an embryologist to be familiar with all the components of a Vitrification system. The goal here would be to do things perfectly without time constraints. The last major step can be broken into mini steps for better acclimatization. He can learn to adjust the coarse knob of the microscope to pick up the embryos after they float owing to change in viscosity in VS, the focusing of the cryotop under the microscope, the placement of the hand, learning the process of washing and finally gaining the ability to load the embryos!
Thirdly, once you have perfected the step, time will come into the picture. Then the use of stop watch and practicing loading in its required time becomes the focus. Initially, the loading time may exceed even two mins. With practice, slowly, you will reduce the necessary time and finally place yourself in the stipulated range of sixty to ninety seconds! This practice can be done with degenerated oocytes.
Fourthly, once you have taken care of the time, the next natural factor will be survival! This is the stage where you will use discarded embryos and actually freeze them as if you are performing an actual procedure. Here, even the ES will come into your practice and you will observe intricate details such as the reaction of the embryos with the media. At this stage, thawing technique will also be incorporated as it too contributes to the overall survival. The results can be recorded and it will give an insight into the overall improvement of the operator.
A comprehensive way of learning Vitrification
Finally, once you are done with survival, it is always recommended to freeze for embryo donation patients as ED cycle harbors the best quality embryos. Blastocysts should be considered more in the initial phase of freezing which survives better than say D2/D3 embryos. This can be furthered by increasing the number of cryo devices progressively corresponding with checking your performance of the previously frozen cryo devices simultaneously.
This comprehensive way of learning Vitrification may take six months overall depending upon the cycles happening in a fertility clinic which certainly affects the availability of gametes and resources(such as expired media for practice). It can help attain better proficiency over the technique and the program can be robustly standardized in a working fertility center itself. Vitrification is a gateway for the embryos to hibernate in an ice age and maybe we can make their journey qualitatively smooth!
As a result of our endeavors further and deeper into various avenues of ART, we take immense pride in announcing the onset of our ART – Clinical Course, for Gynecologist the module of which will be of 3 days. It will be incorporated with a minimal of embryology too.
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