1. Polarization Studies with Malaria Presented by Grace Chojnowski 11 th November 2014 ACS Conference

2. What is Polarization ? © QIMR Berghofer Medical Research Institute | 2 Light waves travel in all directions with different orientations. Polarizing filters just like polarizing sunglasses filter the incoming light so the it then exits in one orientation. Laser light is polarized which means that the emitted light is already travelling in a linear polarized state.

3. © QIMR Berghofer Medical Research Institute | 3 Horizontal PolarizationVertical Polarization

4. © QIMR Berghofer Medical Research Institute | 4 The malaria parasite digests haemoglobin in red blood cells and releases large amounts of heme. This is toxic to cells and the parasites convert this free heme to an insoluble crystalline form called hemazoin. This is referred to as the malaria pigment and is a dark brown colour that can be easily identified using polarizing microscopy. One of the properties of the crystal is its ability to depolarize light. Haemazoin occurs in the blood stage of the parasite life cycle after it has entered the red blood cell. The ability of the parasite to digest and convert the free heme to haemazoin is essential to its survival. Some anti malaria drugs such as chloroquine inhibit this conversion, subsequently killing the parasite. What is Haemazoin ?

5. © QIMR Berghofer Medical Research Institute | 5 Side Scatter Normal RBC Side Scatter Infected RBC with haemazoin

6. © QIMR Berghofer Medical Research Institute | 6 Standard Attune configuration

7. Overview Chapter one… Chapter two… Chapter three… © QIMR Berghofer Medical Research Institute | 7 Attune configuration for polarization detection

8. © QIMR Berghofer Medical Research Institute | 8 488/10 740LP 630/30 580/30 530/30 670/40 555DL P 720DS P 605DS P 505DS P 650DS P ND filter Prism Laser 488/10 740LP 630/30 670/40 50/50 mirror 720DS P 605DS P 650DS P ND filter Prism Laser, Vertical polarization 488/10 Horizontal polarizing filter & 488nm bandpass Vert Pol Horiz Pol Vertical polarizing filter & 488nm bandpass Standard optical layout for laser 1 path. Optical layout for laser 1 path do perform depolarization.

9. © QIMR Berghofer Medical Research Institute | 9 Synthetic Haemazoin no RBCs Infected Red blood cells trophozoites Infected Red Blood Cells ring form. Red Blood Cells only no parasites 405 laser 488 laser Depolarization of different parasite life cycle stages

10. © QIMR Berghofer Medical Research Institute | 10 Michael Rist Nigel Waterhouse Paula Hall Graeme Chapman Grace Chojnowski Tam Hong Nguyen New Position

11. Over to you Chris Peatey……