The portable CASA system

FAQ

General

  • What is CASA - Computer Aided Semen Analysis-, and what is it good for?
  • ~~It reports the quality of the semen sample faster, more accurate, consistent and reliable than that of the result of a human analysis.
  • ~~It is useful at various areas: breeding soundness, genetics production, artificial insemination quality assessment, gene conservation wild or domestic.
  • ~~Computer Aided Semen Analysis: it is a system consisting a microscope combined with a computer to record the behavior of semen samples and analyze it with a software.
  • How does it work?
  • ~~The system records the movement of the sperm cells through the attached microscope and after applying various image processing techniques individually measures the movement properties: VAP (averaged path velocity), linearity (averaged path length / end-to-end distance). From these data it calculates the ratio of cells falling into WHO motility classes, and generates a report of the results.
  • WHO classes are A,B,C,D depending on the field but ONGO reports 3 category: progressive, total and immotile ratios?
  • ~~Breeders are interested in the sample ability to fertilize which mostly described by progressive motility and concentration and they are less interested in further categorizations.

Business questions

  • How to order?
  • ~~You can order via e-mail. After discussing all your needs and business details you will get a pro-forma invoice about your purchase.
  • How long does it take to get the product?
  • ~~Usually it takes 4-6 weeks after the unit has been paid.
  • Warranty?
  • ~~30 days money back guarantee and one year warranty on the ONGO unit.
  • What kind of slides can be used? Where to buy them? How much it costs?
  • ~~ONGO slides are specifically produced and are calibrated to be used with ONGO system. This ensures the accuracy of the results. It can be purchased from our website(??). There are 25 slides in a box and each slide has two chambers so 50 samples can be analyzed with one box.

ONGO specific questions

  • What kind of semen species can be analyzed by ONGO? All is the same?
  • ~~It is available for boar, bull, stallion, dog, poultry. The semen are different from species to species in various features like size, shape, and motility parameters which requires a specific, fine-tuned parameter setup for each species in the software.
  • How to use it? How long is a single measurement?
  • ~~Heat up the slide holder plate, possibly with the slide placed onto the plate to avoid heat shock to the sample.
  • ~~Put one drop  of the semen sample to the entrance of the chamber on the slide.
  • ~~Wait while the capillary sucks in the sample and let it settle down otherwise serious DRIFT, a movement caused by the capillary effect will be present and either lead to false results or ONGO will refuse measurement (quality management).
  • ~~Follow the on-screen instructions: with the help of a live view of the sample adjust focus, illumination as necessary and start the analysis. ONGO will record a video first and process the recording. The result of the analysis is displayed on the screen.
  • ~~You may repeat the measurement on the same sample collecting more data and increase result's accuracy and reliability. After you finished measurement with a given sample click Finish. This action will save a report about all individual measurements made during this cycle along with the recorded videos to the attached pen-drive.
  • ~~To keep the file system healthy, shut down the unit with the pushbutton instead of the main switch. Wait a few seconds after the message appeared on the screen (when the green light turns on at the lid!).
  • Does the data stored on the unit as well?
  • ~~No, and it is not available (yet) from outside only via pen-drive during the measurement. If you forget to attach a properly formatted pen-drive to the unit then all measured data will be lost. However the pen-drive can be attached anytime, the system recognizes it and will save all upcoming data to it.
  • What data is saved to the pen drive?
  • ~~ONGO generates a unique ID number for each measurement. It consists of two numbers.  First is Sample ID, normally this identifies the series of measurements had been taken on one chamber. The second is RepeatID which identifies the repeated measurements taken on the same chamber. SampleID increased after you push the Finish button on the result screen while Repeat ID increased after Repeat button is pushed.
  • ~~ONGO creates a specific folder for each measurement named by the Sample ID. In this folder the recorded videos in mp4 format and the final report in pdf format is saved.
  • When is it safe to remove the pen-drive?
  • ~~Except during video recording and final data saving shown on the screen it is safe to remove pen-drive anytime. Data won't be lost.
  • How long the built in battery lasts?
  • ~~It depends heavily on the external temperature and the usage. On room temperature with the heating always on a fully charged battery can operate around 1 - 1.5 hour or more. If the battery is low a warning message will appear on the screen and ONGO turns the heating off and remind you to plug the power supply in.
  • How long does a single measurement lasts?
  • ~~Startup lasts around 22 seconds,
  • ~~Heating up heavily depends on the temperature of the environment, on the temp of the stage and if the unit is plugged in to power supply. ON battery power it is considerably slower so it's worth to preheat the unit on AC power then use with battery if necessary.
  • ~~Video recording: 12-14 sec. Although this sounds a lot but it consists of the encoding (compression) and saving to the pen drive beside the recording.
  • ~~Analysis: 20-40 sec depending on the concentration. The more cells on the field of view the more to follow and analyze.
  • ~~Saving the data to pen drive when no more measurement is taken on the same sample (pushing the Finish button) takes approx 3 seconds.
  • What kind of slides can be used? Where to buy them? How much it costs?
  • ~~ONGO slides are specifically produced and are calibrated to be used with ONGO system. This ensures the accuracy of the results. It can be purchased through our website. There are 25 slides in a box and each slide has two chambers so 50 samples can be analyzed with one box.

Trouble shooting

  • Pen drive is attached but no data is saved.
  • ~~Pen drive has to be formatted properly. It should have a partition and be FAT32 formatted.
  • ~~The provided pen drive is formatted and ready to use.
  • General insturctions:
  • ~~Do not move the slide during the measurement. Close the lid properly, slightly touching the slide itself. Slide checking is executed during the measurement process, if you move the slide it can fail the checking.
  • ~~Measurement is taken from the middle part of the video. The size of the investigation window depends on the concentration.
  • Result: DRIFT
  • ~~This is a sample problem and the message is shown on the result screen when considerable level of drifting, i.e:  collective unidirectional movement of cells detected. This is caused usually the capillary effect which sucks in the sample into the chamber immediately after you drop the sample to the slide. In this case wait a few seconds while the fluid stops diffusing.
  • ~~Sometimes the fluid does not settle down quickly e.g: when you dropped too much sample to the slide. In this case remove the excessive material with some absorbent cloth.
  • ~~Other reason could be that the unit is held tilted. Place it horizontally.
  • ~~If ONGO detects drift it reports this error and no other data is calculated. The averaged/final result does not include this measurement.
  • Result: LOW_CELL_NUMBER
  • ~~In order to achieve reasonably accurate measurement about the semen quality it is necessary to have a minimum number of cells been analyzed. If the average number of detected cells falls below this limit ONGO reports this error and no other data is calculated. The averaged/final result does not include this measurement.
  • ~~The possible reasons of getting LOW_CELL_NUMBER error can be an overly diluted sample, inhomogeneously distributed sample - a sample problem-, or  out of focus or low contrast videos - a detection problem. Check the sample by moving it slightly during Live View and ensure that real cells can be captured on the screen. The recorded video will look very much like what you can see on the Live View screen.
  • Result: LOW_MOTILITY
  • ~~In order to achieve reasonably accurate measurement about the semen motility it is necessary to have minimum number of motile cells been analyzed. If the average number of detected cells falls below this limit ONGO reports this error and no other data is calculated. The averaged/final result does not include this measurement.
  • ~~The possible reasons of getting LOW_MOTILITY error can be just like LOW_CELL_NUMBER error's  or simply all the cells are dead or non motile because of slow recovery from the cooled status.
  • Result: TOO DENSE
  • ~Every technology has a limit and ONGO is no exception. To be able to properly distinguish the cells from each other it is necessary to set an upper limit of sample concentration. In case of ONGO this is around 100M/ml. Above the limit ONGO reports this error and no other data is calculated. The averaged/final result does not include this measurement and you have to dilute the sample to be analyzed by ONGO.
  • How to calculate the original sample properties after dilution?
  • ~~First of all none of the proportional properties like progressive and total motility ratios are not affected by dilution - theoretically. As long as the dilution and the sample are homogeneous the ratio of fast and slowly moving cells will remain the same.
  • ~~To calculate the concentration you need the measured concentration and the dilution ratio: how many units of diluent volume was added to one unit of sample volume. E.g.:  5 unit diluent to 1 unit sample yield 6 unit volume dilution. This means that the same number of cells which were originally in 1 unit volume now are in 6 volume. In this case you have to multiply the measured concentration by 6 to get back the original/native concentration.

Tips and Tricks

  • Preheat unit on AC power to save battery charge and to speed up reheating the stage.
  • Heat up stage with the slide on the stage. Preheating the slide as well as preheating the stage prevents heat shock to the sperm cells.
  • On the field you may leave the unit in the box which serves as a protecting shell against dirt, moisture or drop.
  • Focus and illumination settings are a little bit off by default. This helps to decide where to start searching the proper focus and illumination.
  • For focus setting if you do not see anything start by pushing "-" until you find the cells. If you repeat the measurement on the same slide the previously set focus will be kept and only minor modification might be necessary.
  • For illumination setting start by pushing "+" to increase the brightness and the contrast of the video. You might need to adjust illumination for the next measurement.

Usage demonstration