Zeiss LSM 710, User Manual

Page 1: ...4 OBSERVATION AT OCULARS 5 STATIF PRESENTATION 6 ACQUIRE ONE OR SEVERAL STAININGS 7 SAMPLING IMAGE RESOLUTION 13 ACQUIRE A Z STACK 14 ACQUIRE A TIME SERIES 16 ACQUIRE A MOSAIC IMAGE 17 ACQUIRE MULTIPL...

Page 2: ...tch on COMPONENT button 5 Light on the fluorescent lamp On the power supply box of Argon laser 6 Check if the button is at the ON position 7 Turn the Power key on I On the small laser box 8 Push the b...

Page 3: ...2011 icon on the desktop 2 Choose START SYSTEM to access to the acquisition menu 3 Zen software is divided in 3 parts the left side is dedicated to acquisition parameters central part is for displayin...

Page 4: ...2 Enter the temperature value that you need and tick the box 3 Open the CO2 bottle 4 Enter the percentage of CO2 and tick the box Warning At the end of your session please untick all boxes and close...

Page 5: ...ht on light off choose the intensity of the transmission lamp 4 Open close the shutter for transmission 5 Light on light off choose the intensity of the fluorescent lamp 6 Open close the shutter for f...

Page 6: ...us knob 1 Set the objective at the lowest position to install your sample 2 Set the objective at its working position 3 No function 4 Switch from the objective of high magnification to a lower one 5 S...

Page 7: ...STAININGS 1 Choose the ACQUISITION tab 2 Tick SHOW ALL TOOLS 3 In the LASER toolbox activate the 561 laser if you need it Power On 4 Open SMART SETUP 5 In DYE select the different dyes and their resp...

Page 8: ...BEST COMPROMISE compromise between speed and reduction of Cross talk Example blue and red channels are acquired simultaneously then the green one is acquired alone LINEAR UNMIXING use spectral capaci...

Page 9: ...e channels with for each sequence 11 If possible select the same dichroic for each sequence in order to speed up the change from one sequence to the other If you don t need to use a dichroic for an ac...

Page 10: ...ach channel 18 You can increase the MASTER GAIN to decrease the laser power but the signal to noise ratio will decrease 19 Don t increase the DIGITAL GAIN except if the GAIN MASTER is really high 20 A...

Page 11: ...e mode is more adapted If it is a living sample the Line mode is preferable Mean method is recommanded 26 The bidirectional mode allows to increase the speed by a factor of two but it must be well cor...

Page 12: ...the image The blue line represents the top of the final image You can move rotate enlarge or reduce the square Then click on LIVE or SNAP 32 Save your configuration 33 Click on SNAP to acquire your i...

Page 13: ...he zoom click on OPTIMAL It is not necessary to have pixels smaller than the optimal resolution oversampling However you can undersample your image in order to increase the acquisition speed and decre...

Page 14: ...s on the center of the sample then click on CENTER If you don t know about your sample thickness 5 Choose the FIRST LAST tab 6 Make the focus on the bottom of your sample then click on SET FIRST 7 Mak...

Page 15: ...me voxels size in all channels 11 To adjust the acquisition sequence in LIGHT PATH go to SWITCH TRACK EVERY and choose Z STACK to change the channel after the acquisition of the entire stack Otherwise...

Page 16: ...L STAININGS section and adjust the parameters in the brightest plane 2 Tick TIME SERIES to make the toolbox appears in the MULTIDIMENTIONAL ACQUISITION menu 3 Choose the number of cycle and the interv...

Page 17: ...center of your mosaic 4 Choose the number of lines and columns 5 Let the OVERLAP at 10 6 Tick BI DIRECTIONAL to acquire faster 7 Tick ONLINE STICHING and let the THRESHOLD at 0 10 Except if you make...

Page 18: ...coordinates on the list and click on REMOVE To remove all click on REMOVE ALL 12 The size of your final mosaic is shown 13 To make a mosaic including different fields of interest without empty zones a...

Page 19: ...rameters of your multiple well plate 6 Click on CALIBRATE to take these parameters into account 7 Select wells of interest with SELECT SELECT ALL buttons or CLEAR ALL 8 Tick OBJECTIVE LOWERED WHEN MOV...

Page 20: ...zoom 3 Native zoom image pixel screen pixel 4 Zoom in Zoom out a zone 5 Select Z position 6 Select time position 7 See a position 8 Tick SINGLE CHANNEL to see channels separately 9 Tick RANGE INDICAT...

Page 21: ...ment images must be acquired in 12 bits So in ACQUISITION MODE toolbox select BIT DEPTH 12bits 2 Adjust acquisition conditions Gain Offset and laser power on the brightest sample 3 Without modifying a...

Page 22: ...ann e mois jour nom d utilisateur folders You can select all acquired images with a right clic on the set of images then SELECT ALL and SAVE You can also delete them by selecting DELETE You can selec...

Page 23: ...If nobody will use the system you can completely switch off 1 Check if you have saved all your data 2 Close ZEN application menu File Exit 3 Switch off the PC select in the toolbar START SHUTDOWN 4 O...