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Oculyze BB 2.0 Handbook

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

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Summary of Contents for Oculyze BB 2.0

  • Page 1 Handbook Analysis of Brewer’s Yeast Oculyze Version 2.0, 2019-06-25...

  • Page 2: Table Of Contents

    Contents Product Information ........1 Initial Setup ........3 Installing the App .
  • Page 3 App shutting down ....... 35 Safety, Compliance, and Disposal Information ... . 37 EU .

  • Page 4: Product Information

    Product Information This handbook is valid for: Product Name Oculyze BB 2.0 Year of Manufacture 2018 - 2019 Model OCU 2.0 App-Version Operating elements: • Focus and fixing knob • Switch for turning device on and off • Android app (for phones or tablets) Accessories delivered: •...
  • Page 5 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/.

  • Page 6: Initial Setup

    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 Fig.

  • Page 7: Sample Preparation

    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 ...

  • Page 8: Staining The Sample

    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.
  • Page 9 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.

  • Page 10: Loading The Sample Chamber

    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.
  • Page 11 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.

  • Page 12: Analysis Of Brewer's Yeast

    Analysis of Brewer’s Yeast Preparing measurement 1. Connect the microscope to the mobile device (Fig. 2. Insert the sample chamber (Fig. 3. Slide the sample chamber so that the left end of the rectangle on the chamber corresponds to the first marker (Fig.
  • Page 13 (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...

  • Page 14: Login

    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;...

  • Page 15: Sync With Server

    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.

  • Page 16: App Settings

    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: •...
  • Page 17 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.

  • Page 18: Recording Images

    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.
  • Page 19 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.
  • Page 20 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.

  • Page 21: Entering Sample Data

    different cells can be recorded. To do so, you must push the chamber to the next marking after each image is taken (Fig. 17). You may also have to use the screw to slightly re-adjust the focus. Should an image be discarded, a replacement image must be recorded.
  • Page 22 • In the “Sample” field, enter a name for the sample. • Date and time of analysis are automatically entered by the app, when the analysis is started. • Under “Dilution in Parts,” you can enter any dilutions used. These will then be used in determining concentration. Thus it is important that the information entered here matches the dilutions performed •...

  • Page 23: Cloud Analysis

    To get the dilution as previously shown in the  handbook, you would have to enter 1 part sample and 99 parts water. “0” parts water means that the sample was not  diluted before staining. By pressing the “Next Step” button, cloud analysis is initiated. Cloud analysis After entering sample data, the images are sent to the cloud for automatic analysis.

  • Page 24: Displaying Results

    Displaying results After cloud analysis has completed, a results screen is loaded (Fig. 20). The calculated concentration appears in cells/ml (green circle) and (if selected) the viability is displayed as a percentage (blue circle). You can access the Pitch Rate Calculator via the click of the arrow, which appears in the result circle for the concentration.
  • Page 25 stacked on top show the distribution of dead cells. Overall larger cells indicate an older yeast  population, as the cells grow larger with each budding (splitting). To review the image analysis, click on one of the results to enlarge the image (Fig.
  • Page 26 For the viability analysis 4 types of cells are differentiated: • green cells: Live cells • violet / blue cells: Dead cells • yellow-black border: Dividing mother cell or large daughter cell (ignored for viability calculation) • only yellow border: Small daughter cell (ignored for viability and concentration calculations For the concentration analysis 3 types of cells are differentiated:...
  • Page 27 After measurement is complete, the sample chamber must always be cleaned (see  Maintenance). Should the yeast sample dry up in the chamber, the chamber may be permanently damaged.

  • Page 28: Pitch Rate Calculator

    Pitch Rate Calculator The Pitch Rate Calculator (Fig. 22) is accessible through the gray arrow, that is shown in the concentration results screen. The concentration (and viability) measurements are used to calculate the amount of yeast needed for a new brewing process. When concentration and viability have been determined, only the living yeast cells are taken into account for the calculation.

  • Page 29: Previous Results

    As a result, the amount of yeast to be used is expressed in liters. If more than 10% of the total volume is accounted for by the yeast, a warning appears and the total volume in hectoliters is also displayed. Previous results The result history can be selected from the starting screen (Fig.
  • Page 30 previous results (Fig. 24). Figure 24. Home screen for the manual analysis Counting the Cells After the image is captured (see Recording images), an additional screen will appear with a counting grid that is shown on top of the captured image. This is meant to mimic a hemocytometer/Thoma Chamber counting grid, to help guide you with your manual counting.
  • Page 31 ¬ However, you should count the cells that touch the white part of the circle, even if they are located outside of it. • In order to avoid double counting, do not count cells for each box if they touch the lower or right grid lines. Results Screen for Manual Counting Similar to the results screen (see Displaying...

  • Page 32: Maintenance

    Maintenance Cleaning the sample chamber In order to guarantee optimal analysis, the sample chamber must be cleaned and dried after each use. To do so, fill the included syringe with distilled water (Fig. 25) and then use it to rinse out the chamber (Fig.
  • Page 33 Lastly, the chamber should be dried using the included bellows. To do this, rest the bellows against one of the sample chamber’s openings and then press the bellows to blow air through the chamber (Fig. 27). To prevent water flowing back into the chamber, it may be helpful to insert tissue paper into the other opening of the sample chamber in order to soak up the expelled water (Fig.

  • Page 34: Cleaning The Device

    Cleaning the device In order to guarantee ideal image quality, the optics on the top of the device should be regularly cleaned (Fig. 29). Figure 29. Cleaning the optics from the top...

  • Page 35: Troubleshooting

    Troubleshooting Problems during Sample Preparation Sample doesn’t flow in the chamber Especially with new (unused) sample chambers you may encounter an issue with the sample not being completely pulled through by capillary forces. In this case, it is usually helpful to lightly tap the chamber on a fixed surface (Fig.

  • Page 36: Problems During Analysis

    Please clean the device as described under Maintenance. Login doesn’t work Make sure both your username and your password are correct. Should the problem persist, please contact Oculyze at support@oculyze.de. Please also review your emails in order to ensure ...
  • Page 37 1 minute after loading. • The sample chamber was not properly cleaned after a previous analysis. Please make sure none of these apply and repeat the analysis. If you continue to encounter problems, please contact Oculyze at support@oculyze.de.

  • Page 38: App Shutting Down

    App shutting down Send a bug report Should the app crash, a bug report screen is automatically opened (Fig. 32). Figure 32. Sending a bug report Here, you can describe what happened before the app crashed so that the error can be better identified. Should something be displayed incorrectly you also have the option of sending a screenshot of the app along with the bug report.
  • Page 39 The simplest way to do this is through the Play Store. Simply start the Play Store app, enter “Oculyze” in the search bar, select the Oculyze app in the list and press the “Uninstall” button. After a successful uninstall, you can reinstall the app from the same screen by pressing the “Install”...

  • Page 40: Safety, Compliance, And Disposal Information

    This device is subject to the RoHS and WEEE Directives (WEEE- Reg.-Nr.: DE 31235761). Do not dispose of this device in the regular trash; send it back to Oculyze GmbH for disposal. This device complies with Part 15 of the FCC Rules. Operation is subject to the following two conditions: 1.

  • Page 41: Appendix

    Appendix • Methylene blue solution recipe. • Methylene violet solution recipe. • Safety information sheet for methylene blue (Sigma) • Safety information sheet for methylene violet (Sigma) • Safety information sheet for glycin (Sigma) • Safety information sheet for NaOH (Merck)
  • Page 42 Recipe: Alkaline Methylene Blue 0.01% Alkaline methylene blue solution – 100 ml Chemicals: Methylene blue (Sigma) • 0.1M Glycine buffer (pH 10.6) • Distilled water • Procedure: Dissolve 0.1g methylene blue in 100 ml distilled water (0.1% MB stock solution) •...
  • Page 43 Recipe: Alkaline Methylene Violet 0.01% Alkaline methylene violet solution – 100 ml Chemicals: Methylene violet 3RAX (Roth) • 0.1M Glycine buffer (pH 10.6) • Distilled water • Procedure: Dissolve 0.1g methylene violet in 100 ml distilled water (0.1% MB stock solution) •...
  • Page 44 SIGMA-ALDRICH sigma-aldrich.com SAFETY DATA SHEET according to Regulation (EC) No. 453/2010 Version 5.2 Revision Date 11.08.2015 Print Date 09.02.2016 GENERIC EU MSDS - NO COUNTRY SPECIFIC DATA - NO OEL DATA SECTION 1: Identification of the substance/mixture and of the company/undertaking Product identifiers Product name Methylene blue...
  • Page 45 Statements Other hazards This substance/mixture contains no components considered to be either persistent, bioaccumulative and toxic (PBT), or very persistent and very bioaccumulative (vPvB) at levels of 0.1% or higher. SECTION 3: Composition/information on ingredients Substances Synonyms Tetramethylthionine chloride 3,7-bis(Dimethylamino)phenazathionium chloride Basic Blue 9 Formula C 16 H 18 ClN 3 S ·...
  • Page 46 Advice for firefighters Wear self-contained breathing apparatus for firefighting if necessary. Further information No data available SECTION 6: Accidental release measures Personal precautions, protective equipment and emergency procedures Use personal protective equipment. Avoid dust formation. Avoid breathing vapours, mist or gas. Ensure adequate ventilation.
  • Page 47 Material tested:Dermatril® (KCL 740 / Aldrich Z677272, Size M) Splash contact Material: Nitrile rubber Minimum layer thickness: 0,11 mm Break through time: 480 min Material tested:Dermatril® (KCL 740 / Aldrich Z677272, Size M) data source: KCL GmbH, D-36124 Eichenzell, phone +49 (0)6659 87300, e-mail sales@kcl.de, test method: EN374 If used in solution, or mixed with other substances, and under conditions which differ from EN 374, contact the supplier of the CE approved gloves.
  • Page 48 q) Decomposition No data available temperature Viscosity No data available Explosive properties No data available Oxidizing properties No data available Other safety information No data available SECTION 10: Stability and reactivity 10.1 Reactivity No data available 10.2 Chemical stability Stable under recommended storage conditions. 10.3 Possibility of hazardous reactions No data available...
  • Page 49 Additional Information RTECS: SP5740000 Absorption into the body leads to the formation of methemoglobin which in sufficient concentration causes cyanosis. Onset may be delayed 2 to 4 hours or longer., Vomiting, Diarrhoea, To the best of our knowledge, the chemical, physical, and toxicological properties have not been thoroughly investigated. SECTION 12: Ecological information 12.1 Toxicity...
  • Page 50 15.2 Chemical Safety Assessment For this product a chemical safety assessment was not carried out SECTION 16: Other information Full text of H-Statements referred to under sections 2 and 3. H302 Harmful if swallowed. Further information Copyright 2015 Sigma-Aldrich Co. LLC. License granted to make unlimited paper copies for internal use only.
  • Page 51 SIGMA-ALDRICH sigma-aldrich.com SAFETY DATA SHEET according to Regulation (EC) No. 1907/2006 Version 5.0 Revision Date 11.12.2012 Print Date 15.01.2018 GENERIC EU MSDS - NO COUNTRY SPECIFIC DATA - NO OEL DATA IDENTIFICATION OF THE SUBSTANCE/MIXTURE AND OF THE COMPANY/UNDERTAKING Product identifiers Product name Methylene Violet 3RAX Product Number...
  • Page 52 In case of skin contact Wash off with soap and plenty of water. In case of eye contact Flush eyes with water as a precaution. If swallowed Never give anything by mouth to an unconscious person. Rinse mouth with water. Most important symptoms and effects, both acute and delayed To the best of our knowledge, the chemical, physical, and toxicological properties have not been thoroughly investigated.
  • Page 53 Personal protective equipment Eye/face protection Use equipment for eye protection tested and approved under appropriate government standards such as NIOSH (US) or EN 166(EU). Skin protection Handle with gloves. Gloves must be inspected prior to use. Use proper glove removal technique (without touching glove's outer surface) to avoid skin contact with this product.
  • Page 54 Oxidizing properties no data available Other safety information no data available STABILITY AND REACTIVITY 10.1 Reactivity no data available 10.2 Chemical stability no data available 10.3 Possibility of hazardous reactions no data available 10.4 Conditions to avoid no data available 10.5 Incompatible materials Strong oxidizing agents...
  • Page 55 Additional Information RTECS: Not available ECOLOGICAL INFORMATION 12.1 Toxicity no data available 12.2 Persistence and degradability no data available 12.3 Bioaccumulative potential no data available 12.4 Mobility in soil no data available 12.5 Results of PBT and vPvB assessment no data available 12.6 Other adverse effects no data available...
  • Page 56 The above information is believed to be correct but does not purport to be all inclusive and shall be used only as a guide. The information in this document is based on the present state of our knowledge and is applicable to the product with regard to appropriate safety precautions. It does not represent any guarantee of the properties of the product.
  • Page 57 SIGMA-ALDRICH sigma-aldrich.com SAFETY DATA SHEET according to Regulation (EC) No. 1907/2006 Version 6.2 Revision Date 10.12.2014 Print Date 08.12.2016 GENERIC EU MSDS - NO COUNTRY SPECIFIC DATA - NO OEL DATA SECTION 1: Identification of the substance/mixture and of the company/undertaking Product identifiers Product name Glycine...
  • Page 58 No components need to be disclosed according to the applicable regulations. SECTION 4: First aid measures Description of first aid measures If inhaled If breathed in, move person into fresh air. If not breathing, give artificial respiration. In case of skin contact Wash off with soap and plenty of water.
  • Page 59 Specific end use(s) Apart from the uses mentioned in section 1.2 no other specific uses are stipulated SECTION 8: Exposure controls/personal protection Control parameters Components with workplace control parameters Exposure controls Appropriate engineering controls General industrial hygiene practice. Personal protective equipment Eye/face protection Use equipment for eye protection tested and approved under appropriate government standards such as NIOSH (US) or EN 166(EU).
  • Page 60 Colour: white b) Odour No data available Odour Threshold No data available d) pH No data available e) Melting point/freezing Melting point/range: 240 °C point Initial boiling point and No data available boiling range g) Flash point No data available h) Evaporation rate No data available Flammability (solid, gas) No data available...
  • Page 61 SECTION 11: Toxicological information 11.1 Information on toxicological effects Acute toxicity LD50 Oral - Rat - 7.930 mg/kg Skin corrosion/irritation No data available Serious eye damage/eye irritation No data available Respiratory or skin sensitisation No data available Germ cell mutagenicity No data available Carcinogenicity IARC:...
  • Page 62 SECTION 13: Disposal considerations 13.1 Waste treatment methods Product Offer surplus and non-recyclable solutions to a licensed disposal company. Contaminated packaging Dispose of as unused product. SECTION 14: Transport information 14.1 UN number ADR/RID: - IMDG: - IATA: - 14.2 UN proper shipping name ADR/RID: Not dangerous goods IMDG:...
  • Page 63 SAFETY DATA SHEET according to Regulation (EC) No. 1907/2006 Revision Date 28.05.2015 Version 5.1 SECTION 1. Identification of the substance/mixture and of the company/undertaking 1.1 Product identifier Catalogue No. 109137 Product name Sodium hydroxide solution c(NaOH) = 1 mol/l (1 N) Titripur® Reag. Ph Eur,Reag.
  • Page 64 SAFETY DATA SHEET according to Regulation (EC) No. 1907/2006 Catalogue No. 109137 Product name Sodium hydroxide solution c(NaOH) = 1 mol/l (1 N) Titripur® Reag. Ph Eur,Reag. Precautionary statements Prevention P280 Wear protective gloves/ protective clothing/ eye protection/ face protection. Response P301 + P330 + P331 IF SWALLOWED: Rinse mouth.
  • Page 65 SAFETY DATA SHEET according to Regulation (EC) No. 1907/2006 Catalogue No. 109137 Product name Sodium hydroxide solution c(NaOH) = 1 mol/l (1 N) Titripur® Reag. Ph Eur,Reag. For the full text of the R-phrases mentioned in this Section, see Section 16. SECTION 4.
  • Page 66 SAFETY DATA SHEET according to Regulation (EC) No. 1907/2006 Catalogue No. 109137 Product name Sodium hydroxide solution c(NaOH) = 1 mol/l (1 N) Titripur® Reag. Ph Eur,Reag. Do not empty into drains. 6.3 Methods and materials for containment and cleaning up Cover drains.
  • Page 67 SAFETY DATA SHEET according to Regulation (EC) No. 1907/2006 Catalogue No. 109137 Product name Sodium hydroxide solution c(NaOH) = 1 mol/l (1 N) Titripur® Reag. Ph Eur,Reag. Consumer DNEL, Local effects inhalation 1 mg/m³ longterm Recommended monitoring procedures Methods for measurement of the workplace atmosphere have to correspond to the requirements of norms DIN EN 482 and DIN EN 689.
  • Page 68 SAFETY DATA SHEET according to Regulation (EC) No. 1907/2006 Catalogue No. 109137 Product name Sodium hydroxide solution c(NaOH) = 1 mol/l (1 N) Titripur® Reag. Ph Eur,Reag. properly documented. Environmental exposure controls Do not empty into drains. SECTION 9. Physical and chemical properties 9.1 Information on basic physical and chemical properties Form liquid...
  • Page 69 SAFETY DATA SHEET according to Regulation (EC) No. 1907/2006 Catalogue No. 109137 Product name Sodium hydroxide solution c(NaOH) = 1 mol/l (1 N) Titripur® Reag. Ph Eur,Reag. Explosive properties Not classified as explosive. Oxidizing properties none 9.2 Other data Ignition temperature Not applicable Corrosion May be corrosive to metals.
  • Page 70 SAFETY DATA SHEET according to Regulation (EC) No. 1907/2006 Catalogue No. 109137 Product name Sodium hydroxide solution c(NaOH) = 1 mol/l (1 N) Titripur® Reag. Ph Eur,Reag. Skin irritation Necrosis Mixture causes burns. Eye irritation Mixture causes serious eye damage. Necrosis Risk of blindness! Sensitisation This information is not available.
  • Page 71 SAFETY DATA SHEET according to Regulation (EC) No. 1907/2006 Catalogue No. 109137 Product name Sodium hydroxide solution c(NaOH) = 1 mol/l (1 N) Titripur® Reag. Ph Eur,Reag. SECTION 12. Ecological information Mixture 12.1 Toxicity No information available. 12.2 Persistence and degradability Biodegradability The methods for determining the biological degradability are not applicable to inorganic substances.
  • Page 72 SAFETY DATA SHEET according to Regulation (EC) No. 1907/2006 Catalogue No. 109137 Product name Sodium hydroxide solution c(NaOH) = 1 mol/l (1 N) Titripur® Reag. Ph Eur,Reag. SECTION 13. Disposal considerations Waste treatment methods Waste material must be disposed of in accordance with the national and local regulations. Leave chemicals in original containers.
  • Page 73 SAFETY DATA SHEET according to Regulation (EC) No. 1907/2006 Catalogue No. 109137 Product name Sodium hydroxide solution c(NaOH) = 1 mol/l (1 N) Titripur® Reag. Ph Eur,Reag. SECTION 15. Regulatory information 15.1 Safety, health and environmental regulations/legislation specific for the substance or mixture EU regulations Major Accident Hazard 96/82/EC...
  • Page 74 SAFETY DATA SHEET according to Regulation (EC) No. 1907/2006 Catalogue No. 109137 Product name Sodium hydroxide solution c(NaOH) = 1 mol/l (1 N) Titripur® Reag. Ph Eur,Reag. Labelling Hazard pictograms Signal word Danger Hazard statements H290 May be corrosive to metals. H314 Causes severe skin burns and eye damage.
  • Page 75 SAFETY DATA SHEET – Annex according to Regulation (EC) No. 1907/2006 Catalogue No. 109137 Product name Sodium hydroxide solution c(NaOH) = 1 mol/l (1 N) Titripur® Reag. Ph Eur,Reag. EXPOSURE SCENARIO 1 (Industrial use) 1. Industrial use (Reagent for analysis) Sectors of end-use SU 3 Industrial uses: Uses of substances as such or in preparations at industrial sites...
  • Page 76 SAFETY DATA SHEET – Annex according to Regulation (EC) No. 1907/2006 Catalogue No. 109137 Product name Sodium hydroxide solution c(NaOH) = 1 mol/l (1 N) Titripur® Reag. Ph Eur,Reag. Physical Form (at time of use) Aqueous solution Frequency and duration of use Frequency of use 600 minutes/day Frequency of use...
  • Page 77 SAFETY DATA SHEET – Annex according to Regulation (EC) No. 1907/2006 Catalogue No. 109137 Product name Sodium hydroxide solution c(NaOH) = 1 mol/l (1 N) Titripur® Reag. Ph Eur,Reag. EXPOSURE SCENARIO 2 (Professional use) 1. Professional use (Reagent for analysis) Sectors of end-use SU 22 Professional uses: Public domain (administration, education, entertainment, services,...
  • Page 78 SAFETY DATA SHEET – Annex according to Regulation (EC) No. 1907/2006 Catalogue No. 109137 Product name Sodium hydroxide solution c(NaOH) = 1 mol/l (1 N) Titripur® Reag. Ph Eur,Reag. For (other) local effects risk management measures are based on qualitative risk characterisation. 4.

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