Oculyze BB 2.0 User manual

Handbook
Analysis of Brewer’s Yeast
Oculyze
Version 2.0, 2019-06-25

Contents
Product Information . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1
Initial Setup . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3
Installing the App . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3
Sample Preparation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4
Diluting the sample . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4
Staining the sample . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5
Loading the sample chamber . . . . . . . . . . . . . . . . . . . . . . . . 7
Analysis of Brewer’s Yeast . . . . . . . . . . . . . . . . . . . . . . . . . 9
Preparing measurement . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9
Login . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11
Sync with Server . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12
Starting measurement . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12
App Settings . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13
Recording images . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 15
Entering sample data . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 18
Cloud analysis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 20
Displaying results . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 21
Pitch Rate Calculator . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 25
Previous results . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 26
Manual Counting . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 26
Maintenance . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 29
Cleaning the sample chamber . . . . . . . . . . . . . . . . . . . . . . 29
Cleaning the device . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 31
Troubleshooting . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 32
Problems during Sample Preparation . . . . . . . . . . . . . . . . . 32
Problems during analysis . . . . . . . . . . . . . . . . . . . . . . . . . . 33

App shutting down . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 35
Safety, Compliance, and Disposal Information . . . . . . . . . . . . 37
EU . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 37
US . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 37
Canada . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 37
Appendix . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 38

Product Information
This handbook is valid for:
Product Name Oculyze BB 2.0
Year of Manufacture 2018 - 2019
Model OCU 2.0
App-Version 2.x
Operating elements:
•Focus and fixing knob
•Switch for turning device on and off
•Android app (for phones or tablets)
Accessories delivered:
•2 sample chambers (Ibidi GmbH)
•Bellows
•Measuring cylinder (100ml)
•Cleaning cotton swabs
•Cleaning cloths
•10 disposable Pasteur pipettes (2 x 5)
•10 reaction containers (2ml)
•3 plastic syringes (5ml)
•Methylene violet color solution (10ml)
1

Product functionality:
The Oculyze BB 2.0 can be used with an Android mobile dedvice
and the Oculyze app (available in the Android Play Store),
allowing the user to view yeast cells with 400X magnification. A
current list of compatible devices is available at:
https://www.oculyze.de/en/device-compatibility/. The app sends
any recorded images to a central server, where they are then
analyzed using the Oculyze software. Within seconds, the results
are then displayed on the app.
Operating conditions
•Yeast cell count and viability measurement for brewer’s
yeast
•Measurement to be performed by trained adults only
•This device cannot be discarded in regular trash (within the
EU).
WEEE-Reg.-Nr.: DE 31235761
Work environment
•Dry area (maximum 80% humidity)
•10-40°C
•Altitude maximum 2,000 meters above sea level
•Data connectivity required for automatic analysis (WLAN or
LTE recommended)
2

Initial Setup
Installing the App
To use the Oculyze BB 2.0 the oculyze app has to be installed on
the mobile device. Open the Play Store and search for “oculyze
BB 2.0”. Then select the Oculyze app and press “Install” as seen
in Fig. 1.
Figure 1. Installing the Oculyze app

Note: When starting the Oculyze app for the first
time you will be prompted to grant it permission. It
is important to grant all requested permissions for
the app to operate properly.
3

Sample Preparation
Diluting the sample
Before measurement, the yeast sample may have to be diluted in
order to reach the device’s optimal measurement range. We
recommend dilutions, that result in ~ 5 - 100 cells per image.
The device can also be used for measurements
outside of this range; however, this may reduce the
accuracy of the measurements.
In order for the yeast sample to be diluted, an appropriate
amount of water must be added. For example, in order to reduce
yeast concentration to one-hundredth, 1 part sample must be
diluted with 99 parts water (1:100 dilution). The included Pasteur
pipettes and measuring cylinder can be used for this purpose
(Fig. 2).
With the container filled up to its upper edge, the
Pasteur pipette took in exactly 1 ml.
The diluted sample (Fig. 3) can then be used for measurement or
staining.
4

Figure 2. Dilution of 1:100
Figure 3. Diluted yeast sample
Staining the sample
In order to determine the viability of the yeast, it must be stained
with methylene violet or methylene blue.
Methylene violet is recommended as the best
method for brewer’s yeast.

Caution: Methylene violet and blue solution are
poisonous when ingested.
Please consult the safety information in the
appendix.
5

In order to stain the sample, the sample must be mixed with the
coloring agent. 10ml of prepared methylene blue and violet
coloring solution are included with the delivery. We recommend
the use of alkaline methylene blue or violet solution. The recipes
for preparation can be found in the appendix. Most types of yeast
show the best results when mixed at ratios in the area of 1:1 –
1:5. We recommend a 1:1 mixture for most yeast strains.

A 1:4 mixture, for example, means that there are
5 parts in total, of which one part is coloring agent
and four parts are from the sample. Please do not
confuse this with the previously mentioned
dilution process.
For the mixture, the sample is first added to a reaction container
(Fig. 4), with the coloring agent added afterwards (Fig. 5). After a
staining time of 5 minutes, the measurement process can begin.
Figure 4. Add sample
6

Figure 5. Add methylene violet
Loading the sample chamber
In order to load the sample chamber, use the Pasteur pipette to
collect a small amount of the sample and then inject it into one of
the chamber’s openings (Fig. 6). The sample will be naturally
pulled into the slide via capillary forces (Fig. 7).

Note that you should avoid touching the underside
of the chamber, especially if not wearing gloves.
Oil from skin contact can worsen the captured
images during analysis and will attack the
specialized plastics used, greatly reducing the
chamber lifetime.
Never use a syringe when loading the sample
chamber. This can cause an uneven distribution of
the sample and distort the result of the analysis.
Be careful not to load too much solution into the
chamber - filling the Pasteur pipette to the point
demonstrated in Fig. 5 is sufficient.
7

Should the solution not enter the chamber on its
own, tap the chamber lightly to encourage the
liquid flow (see Troubleshooting)
The filled chamber should then rest for at least 1-2 minutes so
that the yeast sediments / setles.
Figure 6. Loading the sample into the chamber
Figure 7. The sample is pulled into the chamber via capillary
forces
8

Analysis of Brewer’s Yeast
Preparing measurement
1. Connect the microscope to the mobile device (Fig. 8).
2. Insert the sample chamber (Fig. 9).
3. Slide the sample chamber so that the left end of the
rectangle on the chamber corresponds to the first marker
(Fig. 10).
4. Turn on your mobile phone and start the app.
Figure 8. Attach microscope
Figure 9. Insert sample chamber & screw it into place
9

Figure 10. Bring the sample chamber into position. Position 3 is
shown in the image

Note:The red status light is on, when when the
device is turned on and lighting is active
(automatic after connecting to a mobile device). To
conserve battery power on your mobile device, the
Oculyze BB 2.0 turns itself off automatically after
15 minutes of use. The indicator light will blink
briefly before this happens; by pressing the button
again, the device will remain powered on for an
additional 15 minutes
10

Login
After starting the app, you will be led to the login screen (Fig. 11).
In the upper right hand corner of the screen, you will see
operating elements, which will lead you to the online version of
this handbook (“i”) and will also allow you to report errors in the
app (Bug icon; also see Troubleshooting).
On the login screen, you must enter your appropriate login data
in the “Email” and “Password” fields. Then press the “Login”
button to proceed.
Figure 11. App login screen

If you wish to change or if you forgot your
password, press the "I need a password reset" at
the bottom of this screen. A link to a reset form for
your password will be send to your user email.
Should you encounter problems during login,
please consult the Troubleshooting section.
11

Sync with Server
Immediately after logging in, the Oculyze app will automatically
sync your data with the server. If you are logging in on a new
device with a preexisting account your previously generated
measurements will be pulled from the server, this process could
take up to 3 minutes.
Having your data secured in the cloud makes sure that even in
the even of a loss of damage to your device, it is always kept safe
for you.
Starting measurement
After login, you will see the start page. Here, you can select
which analysis is to be performed (Fig. 12). Simply press the
appropriate corresponding button to begin analysis. To do so, you
can choose between the following options:
•"Viability & Concentration" – Living-dead analysis and
concentration measurement
•"Only Concentration" – Concentration measurement
•"History" – Results of previous analyses
You must use a stained sample for living-dead
analysis!
On this screen, you can check the version of the
app in the upper left corner.
12

Figure 12. Method selection
At the top right of the screen there is also a button which gives
you access to the app settings screen.
App Settings
The settings screen (Fig. 13) lets you adjust various parameters
of the app:
•Switch between German, English or Chinese language
•Change the amount of images per measurement
(5 recommended)
•Change the height of the sample-chamber in use

The selected chamber height has to correspond to
the chamber in use! If you are unsure, you can find
the correct height etched into your chamber (either
0.1, 0.2 or 0.8)
•The on/off switch for the sharpness indicator which is visible
during the image taking process on the bottom left hand
13

side (right next to the white “take photo” button)

When the sharpness indicator is on, an indicated
number in a circle will be shown at the bottom left
hand side of the screen. This shows in real time
how high the sharpness level of that particular
image is. The sharpness indicator values will
change through the turning of the focus knob.
When you reach the highest possible value, the
image has reached an optimal level of sharpness
and a photo can be taken. Since the sharpness
indicator is influenced by the number of cells within
each image, it’s important to reach the highest
value indicated for each image, not based on a
specific value each time. (i.e. one image might
have the highest value on the sharpness indicator
at 2300 and another only 500, use the highest
value for each image).
•Switch between Methylene Violet (MB) and Methylene Blue
(MV) analysis

You can choose between Methylene Violet or
Methylene Blue as the staining agent for viability
analysis. This is selectable on the start screen –
you will see a blue or violet pipette icon – select
the one you wish to use. The results of the viability
measurements are also displayed in the
corresponding color.
•On/Off Switch for Manual Counting
14


After you have captured the image, a separate
window will automatically open, where you can
count the cells by hand. You will repeat this step
for each image. Automatic analysis will not be
conducted. All results will be saved in the Cloud,
just as they would with the automated analysis.
Please refer to Manual Counting to see how the
process differs from the automated analysis.
•View "Legal Notes" or "Privacy Policy"
•Start the tutorial
•Logout from the App
Figure 13. Settings Screen
Recording images
After the measurement has begun, a live image of the front
camera will be displayed. If this image is blurry, the sample must
be focused by turning the knob on the hardware (Fig. 14). You
should see a clear image of the cells in the middle of the screen
before taking a picture (Fig. 15).
15


Adjusting the focusing knob to apply light pressure
to the sample chamber is enough to bring the cells
into focus to see them clearly. When using a new
device a little bit more pressure is needed, since
the pads for the chamber fixing compress with
repeated usage. Screwing it on too tightly will lead
to blurry pictures and possibly damage the sample
chamber or even the device itself.
Figure 14. Focusing the device
Figure 15. Cells in focus (center) in the app
16


The screenshots in this handbook always show the
measurement of a methylene blue stained sample
on which living-dead analysis is being performed.
The dead cells are stained blue, while the living
cells appear transparent.
Pictures can be taken using the green and white button. The
counter above the button shows how many pictures have been
taken.
After the picture is taken, that picture will be displayed on your
screen. Using the “Delete” and “Keep” buttons, the user may
decide whether to dispose of the picture or to keep it for analysis
(Fig. 16). The app also performs an initial automated quality
check of the image. If the image contains far too much yeast or is
significantly blurry, a message will be displayed, and the “Keep”
button will be greyed out. In this case, you must delete the image
using the “Delete” button.
Figure 16. Options for an image taken
A total of 5 images must be taken per analysis. The sample
chamber must be moved somewhat between each image so that
17