Marsico Hall Microscopy Facility

Marsico Hall Microscopy Fellowship (MHMF.ORG)

Olympus Apexview1000 - Bench Top Microscope

Location: Marsico Hall 7229N

Overview:

The Apexview 100 is a bench top microscope for imaging live or fixed samples.
The microscope is an inverted format optimized for live cells on slides, chamber slides, 35 mm petri dishes, microtitre plates, small flasks.
The 4x, 10x, 20x and 40x objectives have sufficient working distance to image through more than 1 mm of plastic (standard culture ware). A 60x 1.4 NA oil immersion objective will image through cover slips only ideally of #1.5 thickness (~0.17 mm).
Tiling, z-stacks & time lapse can be carried out with transmitted light, fluorescence. Phase contrast is available with 10x and 20x objectives. Digitally enabled gradient contrast possible with all objectives.

The slide scanner system is for use by researchers in the Marsico Lung Institute and their colleagues and collaborators

Notices:

Ultimate permission to use the system is at the discretion of the MLI/CF Center Director

All users are required to receive instruction from Michael before using the system

All users should log in with their own account (local)

OlyVIA 4.2.1 free viewer will install on Windows

Apexview100 booking Calendar:

Apexview 100 version 4.4.1 Notes:

Load slides/dishes/flask/microtitre plate into appropriate tray

Load tray into right side to scanner. Note orientation of notch.

Points to note:

Cleaning off immersion oil:
- The 60x 1.4 NA objective needs to be cleaned after use. Use only lens tissue (Not kim wipes etc.)
- After 60x oil immersion scanning is done the tray will need to be removed (see Exchange Trays) and the oil cleaned off before more scanning can be carried out with other objectives
If you encounter problems please note error code or messages. Take a screen shot or cell phone picture - briefly note what you were doing

Mapping Drive Letters to servers (connecting to a server, e.g. CF-shared or CF-groups or MHI-groups):

See http://microscopy.unc.edu/shares on how to connect to e.g.:
- For CF-shared, CF-groups, \\som-fs.med.unc.edu\cf\shared, or \\ad.unc.edu\med\cf\shared
- Mac access use smb://ad.unc.edu/med/cf
- Shared/groups/users for other departments and divisions will require file paths (unc) suited for your division/department

Important Considerations:

Focus upper limit is deliberately set high so it is possible to crash into the sample with 60x or 40x objectives.
Blot the 60x objective with lens tissue when finished scanning with it. This will help prevent oil from dripping into the system or onto other slides.

General Considerations:

Store scans on the D: drive. In an emergency data may be stored on C:. Avoid using the C: drive and including desktop, pictures, etc.
Olympus states not to scan directly to mapped network drives. Scanning to mapped drive letters slows down scanning. Data corruption is possible when the data stream is disrupted by network traffic jams or transient disconnections.
Connecting to a server (mapping to a driver letter) e.g. CF Shared
It is best to scan to D: and copy files later to servers.

Covid-19 Mitigations:

Quick guide & detailed considerations - hygiene tips for microscopes

The System:

Holder for 3x slides (3" x 1"), 3x 35 mm petri dishes, microtitre plates, small flasks
Transmitted light - LED RGB lamp
Fluorescence - 5 channel DAPI like, FITC like, TexasRed like, CY5 like & CY7 like
LED Fluorescence light - X-Cite Xylis II XT730S

Position	Channel	Ex (nm)	BS (nm)	Em (nm)	Cube	Similar fluorophores
	transmitted				none
	DAPI	350/50x (325-375)	400	460/50m (435-485)		Hoechst
	GFP	470/40x (450-490)	495	525/50m (500-550)		FITC, Alexa 488, CY2, Opal 520
	TRITC	554/23 (542-566)	573	609/54 (582-636)	Use mCherry	Opal 570, CY3, Alexa 555, Rhodamine
	TxRed	560/40x (540-580)	585	630/75M (597-663)		Alexa 568, Alexa 594, Opal 620, mCherry
	Cy5	620/60 (590-650)	660	700/75 (662-738)		Alexa 633/647, Opal 650, Draq5, CY5
	Cy7	710/75x (721-749)	760	809/90m (763-854)		CY7, Alexa 750, Opal 750
	GC				Gradient contrast
	PH				phase contrast	limited objectives

CMOS camera fluorescence and transmitted overlays - Orca Fusion sCMOS camera
RGB Color camera for transmitted light/chromogenic sample imaging - Evident DP72
Macro camera for sample overview and label reader. Transmitted only
Objectives:

Objectives

Mag. NA type WD corrections cover slip Immersion Pixel size
@zoom=1
monochrome/Orca-Fusion Pixel size
@zoom=1
color camera

2x 0.06 PLN * 5.8 mm - #1.5 dry

4x 0.13 UPLFLN4X-2 * 13 mm - #1.5 dry

10x 0.30 UPLFLN10X2PH-2 10 mm - #1.5 dry

20x 0.45 UCPLFLN20XPH-1 ~7 mm holder thickness 0 - 2 mm dry

40x holder thickness motorised 0 - 2 mm dry

60x 1.42 UPLXAPO O 0.15 mm -
#1.5
oil^{1, 2}

¹Standard immersion (carbon) oil (RI=1.515)
²For 60x immersion oil must be added when prompted by the software. Also oil should be removed with lens tissue when prompted.
* Not suitable for CY5 or CY7

Objectives
Mag.	NA	type	WD	corrections	cover slip	Immersion	Pixel size @zoom=1 monochrome/Orca-Fusion	Pixel size @zoom=1 color camera
2x	0.06	PLN *	5.8 mm	-	#1.5	dry
4x	0.13	UPLFLN4X-2 *	13 mm	-	#1.5	dry
10x	0.30	UPLFLN10X2PH-2	10 mm	-	#1.5	dry
20x	0.45	UCPLFLN20XPH-1	~7 mm	holder thickness	0 - 2 mm	dry
40x				holder thickness motorised	0 - 2 mm	dry
60x	1.42	UPLXAPO O	0.15 mm	-	#1.5	oil^{1, 2}

Viewing Software:

OlyVIA is the fastest and most convenient way to review scans. Limited processing capabilities. Only for Windows
- OlyVIA 4.2.1 for Windows 11 & earlier
  - Available at D:\_OlyVIA, also at D:\_OlyVIA on VS200, Visiopharm, Fiji computers. (CF) S:\shared\vs200\_OlyVIA
  - Copy installation files to C: drive (Best not to install from a drive letter mapped to a network drive). Administrator privileges are required to install it on your computer.
or processing, quantitation and analysis on Windows, macOS, Linux - Ask Michael for tips
VisioPharm pathology software
QuPath - pathology software available at https://qupath.github.io/
FIJI/ImageJ - can get from https://fiji.sc

Using the system:

Quick startup/shut down guide - ask Michael for a draft copy
Powering up:
- Power on at bottom right of smaller box - green light will be lit
- LED light source - X-Cite Xylis II
- Fusion B/W camera
- Note: fluorescence light & B/W camera may be need to be on even if transmitted light only imaging is being carries out

Last Updated: 2026-06-11