NSF BioPACIFIC MIP Wiki

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====== Sample Preparation ======

{{:workflow_diagram_planning_whiteboard_in_green_yellow_modern_professional_style_1_.png?600 |}}

===== Grids =====


|**Thickness:** 25 μm  \\ **Diameter:** 3 mm \\ Film on “dull” side\\ “Shiny” side has the metal only \\                       |

  * Ultrathin Carbon Film on a Lacey Carbon Support Film
  * https://www.tedpella.com/Support_Films_html/Support_Films_and_Substrates_Overview.aspx
  * 01824 	Ultrathin Carbon Film on Lacey Carbon Support Film, 400 mesh, Copper
  * 160 	PELCO®️ TEM Grid Storage Box  

{{ :tem_grids.jpg?600 |}}                         



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===== Basic Shake and Bake =====

Put a small amount of powder in Eppendorf tub (enough to coat the bottom). Put one grid into the tube. Close the tube and shake for 1 minute. Open the tube and remove the grid. The grid should have enough material on it and is ready to be imaged in the TEM. 
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===== Instructions for Nanoparticle Dropcast TEM sample preparation =====

**Things you will need:**

	* Ethanol (or water, IPA, or dimethyl carbonate)\\
	* A small metal scooper\\
	* A sonicator\\
	* Several small vials that are centrifuge compatible\\
	* A centrifuge\\
	* Nanoparticles\\
	* UltraThin 3 nm Carbon Grids\\
	* Glass slide\\
**1.** Check the MSDS for mixture with ethanol (make sure the nanoparticles are NOT pyrophoric)\\

**2.** Use the small metal scooper to extract about ½-1 cms worth of nanoparticles\\

**3.** Place the nanoparticles in a small vial\\

**4.** Fill the small vial with ethanol up to the 1 cm below the rim\\

**5.** Sonicate the filled small vial for 10 minutes on high power (make sure to find an anti-node)\\

**6.** Centrifuge for 20 minute (make sure to balance the centrifuge)\\

**7.** Use a pipette to collect several uL of fluid just above the concentrated part at the bottom of the vial. See the arrow below . The pipette should just touch above dark part\\
{{ :vial2.png?200 |}}

**8.** Deposit 1 uL of fluid onto the dull (not shiny) side of the grid\\

**9.** Let the grid dry on a glass slide\\

**10.** Vacuum bake at 50-400 degree C for several hours if contamination is found of the surface\\

**11.** Plasma clean as needed\\